Dissertations / Theses: 'Gel electrophoresis'

1

Hosseini, Seyed Homayoun. "Temperature gradient gel electrophoresis development and application." Diss., Georgia Institute of Technology, 1994. http://hdl.handle.net/1853/25614.

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2

杜光旭 and Kwong-yuk To. "Electrophoretic behaviour of polystyrene microspheres in agarose gels." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1993. http://hub.hku.hk/bib/B31233247.

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3

To, Kwong-yuk. "Electrophoretic behaviour of polystyrene microspheres in agarose gels /." [Hong Kong] : University of Hong Kong, 1993. http://sunzi.lib.hku.hk/hkuto/record.jsp?B13465430.

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4

Askarian, Nameghi Shahnaz. "Genotyping Escherichia coli isolates by Pulsed-Field Gel Electrophoresis." Thesis, Södertörn University College, School of Life Sciences, 2007. http://urn.kb.se/resolve?urn=urn:nbn:se:sh:diva-1411.

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Transmission of bacterial strains between patients is a serious problem in hospitals and with the increasing rate of antibiotic resistance the problem has farther escalated. Enterobacteriaceae produced extended-spectrum beta-lactamses (ESBLs), especially Escherichia coli (E-coli), are increasingly important nosocomial pathogens (7, 8). These bacteria are often multiple resistant and are responsible for many intestinal infections and urinary tract infections (2, 5). With the more frequent use of invasive devices in hospital care, these types of nosocomial infections have increased, particularly in seriously ill patients.

In order to diminish transmission of bacterial strains between patients and to study the epidemiology of these bacteria, it is of great importance to develop rapid and specific methods to be able to subtype on strain-level, i.e. to create a fingerprint of the isolates. The method may be based on phenotypic or genotypic characteristics of the microorganism. Any typing method must have high reproducibility and discrimination power to differentiate unrelated strains and also to demonstrate relationship of organisms deriving from the same source. In the present project, a Pulsed-Field Gel Electrophoresis (PFGE) assay for genotyping clinical E. coli isolates was used. PFGE can be used as a genotyping tool and is widely used to type bacteria and trace nosocomial infection. However, the method is time-consuming and relatively expensive in compare with other methods like PCR. In this study, a total of 93 strains were collected. The study was aimed to investigate the genotypes of the collected isolates and to identify and potential the outbreak strains.

The isolates investigated were genotypically diverse shown by a variety of PFGE banding patterns. However, clusters of closely related isolates involved in outbreaks were also identified.

In conclusion, when analyzing a large number of strains, a combination of a rapid phenotyping or genotyping method and a powerful genotyping method like PFGE would be an appropriate strategy for studying clonal relationship among isolates e.g. for detecting cross-transmission of nosocomial pathogens.

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Chan, Hong-Lin. "A 2D-difference gel electrophoresis strategy for redox proteomics." Thesis, University College London (University of London), 2005. http://discovery.ucl.ac.uk/1444604/.

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Post-genomic biomedical science requires quantitative proteomics. In most cases this involves differential protein expression analysis using matched pairs of simultaneously detectable labelling reagents for specific protein amino acids. In this thesis the development and optimisation of a novel cysteine labelling strategy, that is based on the use of iodoacetyl derivatives of Cy3 and Cy5 (ICy3/5) and 2D-difference gel electrophoresis (2D-DIGE) is described. The differentially labelled samples are separated on a single 2D gel and detected by multi-wavelength fluorescence scanning. The method is used to analyse standard proteins and then cell lysates to define the stoichiometry, sensitivity and specificity of this labelling technique. A comparative study of this new proteomic ICy dye protocol with the current NHS-Cy dye labelling system and methods that employ commonly used protein staining methods is described. The method is then used for cysteine labelling of proteins in non-reduced, denatured biological samples allowing accurate monitoring and sensitive detection of redox-dependent thiol modifications and expression level changes. The method is shown to be compatible with the use of MALDI mass spectrometry to identify proteins by analysis of trypsinised ICy labelled peptide digests. Using parallel sample analysis within single gels, the ICy-dye reagents were used to detect redox-, ErbB-2- and growth factor-dependent changes in a human mammary luminal epithelial cell system which was exposed to hydrogen peroxide or to growth factor stimulation. The conventional lysine labelling 2D-DIGE technique was also used in parallel to assess the new ICy labelling strategy for determination of the effects of oxidative stress on protein isoform levels. This study has revealed the identity of proteins involved in the response to oxidative stress and growth factor stimulation in the context of ErbB-2 growth factor receptor over-expression. In addition, this labelling strategy was also used to detect changes in thiol reactivity that follow the UV irradiation of plasma proteins as part of a study designed to evaluate the effect of UV disinfection on plasma product safety for clinical use.

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Stambaugh, Mark P. "Transverse Isotachophoresis Using Polyacrylamide Gel Electrodes." DigitalCommons@CalPoly, 2011. https://digitalcommons.calpoly.edu/theses/505.

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Separation and isolation of a desired analyte from an impure sample solution containing numerous unwanted interfering agents is the first step of nearly every laboratory test performed in medicine and biology. Nucleic acids are often of particular interest to doctors and researchers, and although methods currently exist for their isolation, these procedures are costly in time, man-power, and real-estate. In addition to easing the execution of presently performed tests, mitigation or elimination of these drawbacks would make a large range of currently unperformed tests both practical and feasible. This thesis presents a microfluidics-based approach to the isolation of nucleic acids using transverse isotachophoresis (ITP). A boro-silicate glass chip is used with Poly(Acrylamide) gel electrodes to establish an electric field perpendicular to the direction of sample flow, causing a controlled migration of charged particles. The design and fabrication of the microfluidic chip are addressed, along with the development of a transverse-ITP model which predicts the necessary conditions for the successful separation/concentration of an arbitrary sample. Several proof-of-concept images are provided which demonstrate the effectiveness of transverse ITP using surrogate sample inputs. This thesis proposes a direction for future work which aims toward confirming the model presented and preparing the transverse ITP chip to receive biological samples.

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Xu, Aoshuang. "Development in electrophoresis instrumentation for two-dimensional gel electrophoresis of protein separation and application of capillary electrophoresis in micro-bioanalysis /." [Ames, Iowa : Iowa State University], 2008.

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8

Rye, Morten Beck. "Image segmentation and multivariate analysis in two-dimensional gel electrophoresis." Doctoral thesis, Norwegian University of Science and Technology, Department of Chemistry, 2007. http://urn.kb.se/resolve?urn=urn:nbn:no:ntnu:diva-1744.

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The topic of this thesis is data-analysis on images from two-dimensional electrophoretic gels. Because of the complexity of these images, there are numerous steps and approaches to such an analysis, and no “golden standard” has yet been established on how to produce the desired output. In this thesis focus is put on two essential fields concerning 2D-gel analysis; registration of images by segregation and protein spot identification, and data-analysis on the output of such a registration by multivariate methods. Image segmentation is mainly concerned with the task of identifying individual protein spots in a gel-image. This has generally been the natural starting point of all methods and procedures developed since the introduction of 2D-gels in the mid-seventies, simply because this best reproduces the results created by a human analyst, who manually identify protein-spot entities. The amount of data produced in a 2D-gel experiment can be quite large, especially in multiple gels where the human analyst is dependent on additional statistical data-analytical tools to produce results. Because of the correlated nature of most gel-data, analysis by multivariate methods is natural choice, and are therefore adopted in this thesis. The goal of this thesis is to introduce the above mentioned procedures at different stages in the analysis pipeline where they are not yet fully exploited, rather than to improve already existing algorithms. In this way new insight and ideas on how to handle data from 2D-gel experiments are achieved. The thesis starts with a review of segmentation methodology, and introduces a selected procedure used to identify protein spots throughout. Output from the segmentation is then used to create a multivariate spot-filtering model, which aims to separate protein spots from noise and artefacts often creating problems in 2D-gel analysis. Lately the use of common spot boundaries in multiple gels have been the method of choice when gels are analysed. How such boundaries should be defined is an important subject of discussion, and thus a new method for defining common boundaries based on the individual segmentation of each gel is introduced. Segmentation may be a natural starting point when gels are analysed, but it is not necessarily the most correct. Often the introduction of fixed spot entities introduces restrictions to the data which cause problems at later stages in the analysis. Analysing pixels from multiple gels directly has no such restrictions, and it is shown in this thesis that the output of such an analysis based on multivariate methods can produce very useful results. It can also give insight to the data problematic to achieve with the spot boundary approach. At last in the thesis an improved pixel-based approach is introduced, where a less restricted segmentation is used to reduce and concentrate the amount of data analysed, improving the final output.

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Gauthier, Michel. "Modelling a highly biased random walk: Application to gel electrophoresis." Thesis, University of Ottawa (Canada), 2003. http://hdl.handle.net/10393/26334.

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The drift and diffusion motions of biased particles are commonly studied using random walks on lattices. We present a novel theoretical approach that makes it possible to calculate exact mobilities in the presence of lattice obstacles. Several two-dimensional examples are studied and a particular attention is given to separation techniques and how our model can be used to study such devices. We also broach the problems related to the field-dependence of the diffusion coefficient during random walks, and we present new algorithms that remove these difficulties. We develop new Monte Carlo algorithms that make it possible to study both drift and diffusion processes simultaneously, even in presence of very strong fields. Finally, we present two brief discussions about the addition of curved field lines and viscosity gradients to our lattice models. This work opens the door to a wide range of applications, especially for the study of electrophoretic technologies.

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Höök, Helena. "Campylobacter epidemiology : insights from subtyping by pulsed-field gel electrophoresis /." Uppsala : Dept. of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, 2005. http://epsilon.slu.se/200589.pdf.

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Brown, Andrew S. "Two-dimensional Polyacrylamide Gel Electrophoresis (2D-PAGE) Characterization of Decorin." Youngstown State University / OhioLINK, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1311873768.

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Zheng, Jing. "Coupling gel electrophoresis with mass spectrometry for protein characterization and identification." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape2/PQDD_0011/MQ59912.pdf.

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13

Wang, Luxin Fan Xudong Mustapha Azlin. "Applications of gel electrophoresis in quantum dot conjugates' separation and purification." Diss., Columbia, Mo. : University of Missouri--Columbia, 2009. http://hdl.handle.net/10355/6486.

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Title from PDF of title page (University of Missouri--Columbia, viewed on Feb 19, 2010). The entire thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file; a non-technical public abstract appears in the public.pdf file. Thesis advisor: Dr. Xudong Fan and Dr. Azlin Mustapha. Includes bibliographical references.

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Li, Wenliang. "Magnetic resonance relaxation, diffusion and electrophoresis studies of colloids and polymers." Thesis, University of Bristol, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.336820.

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15

Xin, Yao. "Electrokinetic Modeling of Free Solution Electrophoresis." Digital Archive @ GSU, 2007. http://digitalarchive.gsu.edu/chemistry_diss/18.

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Modeling electrophoresis of peptides, proteins, DNA, blood cells and colloids is based on classical electrokinetic theory. The coupled field equations-Poisson, Navier-Stokes or Brinkman, and ion transport equations are solved numerically to calculate the electrophoretic mobilities. First, free solution electrophoretic mobility expressions are derived for weakly charged rigid bead arrays. Variables include the number of beads (N), their size (radius), charge, distribution (configuration), salt type, and salt concentration. We apply these mobility expressions to rings, rigid rods, and wormlike chain models and then apply the approach to the electrophoretic mobilities and translational diffusion constants of weakly charged peptides. It is shown that our bead model can predict the electrophoretic mobilities accurately. In order to make the method applicable at higher salt concentrations and/or to models consisting of larger sized subunits, account is taken of the finite size of the beads making up the model structure. For highly charged particles, it is also necessary to account for ion relaxation. This ion relaxation effect is accounted for by correcting "unrelaxed" mobilities on the basis of model size and average electrostatic surface, or "zeta" potential. With these corrections our model can be applied to the system with absolute electrophoretic mobilities exceeding approximately 0.20 cm2/kV sec and also models involving larger subunits. This includes bead models of duplex DNA. Along somewhat different lines, we have investigated the electrophoresis of colloidal particles with an inner hard core and an outer diffusive layer ("hairy" particles). An electrokinetic gel layer model of a spherical, highly charged colloid particle developed previously, is extended in several ways. The charge of the particle is assumed to arise from the deprotonation of acidic groups that are uniformly distributed over a portion (or all) of the gel layer. Free energy considerations coupled with Poisson-Boltzmann theory is used to calculate the change of the local pKa of the acidic groups depending on the local electrostatic environment. Based on the modeling of electrophoresis and viscosity, we predict that the thickness of the gel layer decreases as the salt concentration increases. And only the outermost portion of the gel layer is charged.

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Dever, Louisa Violet. "The isolation and characterisation of mutants of Amaranthus edulis lacking key enzymes of the C4 pathway." Thesis, Lancaster University, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.296762.

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Moppett, John Paul. "Prospective analysis of real time minimal residual disease assessment in childhood acute lymphoblastic leukaemia prior to bone marrow transplantation." Thesis, University of Bristol, 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.274837.

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18

Asbury, Charles L. "Manipulation of DNA using nonuniform oscillating electric fields /." Thesis, Connect to this title online; UW restricted, 1999. http://hdl.handle.net/1773/7993.

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19

Toledo, Érica Gil Hernandes de [UNESP]. "Perfil eletroforético de proteínas séricas e urinárias de cães normais e de portadores de insuficiência renal crônica." Universidade Estadual Paulista (UNESP), 2001. http://hdl.handle.net/11449/95978.

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Made available in DSpace on 2014-06-11T19:27:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2001-01-16Bitstream added on 2014-06-13T19:15:42Z : No. of bitstreams: 1 toledo_egh_me_jabo.pdf: 1480955 bytes, checksum: 264147d52e4fdcb0c91216e8ebe11eca (MD5)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Para este estudo foram avaliados 20 cães adultos sadios (10 machos e 10 fêmeas) e 24 cães adultos (16 machos e 8 fêmeas) com insuficiência renal crônica (IRC), com o objetivo de estudar a proteinúria. Foram empregadas técnicas para determinação quantitativa de proteína da urina e para a separação de suas frações por meio de SDS-PAGE. As quantidades de proteínas presentes na urina de cães sadios foram pequenas, mas o estudo dos perfis eletroforéticos mostrou haver diferenças entre machos e fêmeas. Apesar da ampla variação das concentrações de proteínas entre os animais, o grupo dos portadores de IRC apresentou proteinúria intensa. Os resultados dos portadores de IRC foram agrupados de acordo com o sexo ou em função do tipo de lesão renal predominante, para comparação dos eletroforetogramas. As análises revelaram que os perfis eletroforéticos seguem um padrão para os machos e outro pra as fêmeas. Ainda, o padrão o perfil eletroforético observado quando há predomínio de lesões glomerulares difere do observado quando há predomínio de lesões túbulo-intersticiais.
This trial was conducted in 20 adult health dogs (10 male and 10 female) and in 24 adult dogs (16 male and 8 female) in chronic renal failure (CRF), in order to study urinary protein loss. Determination of total urinary proteins and their separation with SDS-PAGE were employed. The results show that CRF dogs had higher proteinuria when compared to controls and the amount of detected protein had large difference among them. In the CRF there are different patterns of protein bands occurrence and distribution according to the patient sex and predominant renal lesion. The difference between protein bands occurrence of male and female already exists in normal dogs.

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Guo, Hong L. "Sieving of spherical particles during gel electrophoresis: A new computer simulation algorithm." Thesis, University of Ottawa (Canada), 1994. http://hdl.handle.net/10393/6741.

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A new computer simulation algorithm is developed to study the behavior of a hard sphere during gel electrophoresis. The electrophoresis mobility and the diffusion coefficients are presented to show the effects of the field intensity, of the gel concentration, and of the randomness of the gel. The results indicate that previous models are not applicable for either the periodic or the random gel, and that the Einstein relation does not hold because the gel molecules affect the dynamics of the hard sphere. Moreover, the randomness of sieving gels lead to a trapping effect where the longitudinal diffusion coefficient is much larger than expected when the gel is dense and the field intensity is non-negligible. This trapping effect predicts a limit to both the field intensity and the gel concentration that one can use for the separation of particles during gel electrophoresis. Our new algorithm thus opens a door to a detailed study of the process of electrophoretic sieving with potential applications in biology and biotechnology.

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Petersson, Nina. "Optimisation of capillary gel electrophoresis method for enhanced separation of mRNA shortmers." Thesis, Uppsala universitet, Institutionen för kemi - BMC, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-351119.

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Advancements in the field of modified messenger RNA (mRNA) has led to new ventures in the pharmaceutical industry. However, new drug products demand new analytical methods to ensure the efficacy and purity of the drug. Capillary gel electrophoresis (CGE) with UV detection shows great potential for separation of mRNA samples due to the equal mass-to-charge ratio of mRNA and the flexible parameters of the CGE methods. This thesis investigates the optimal parameters of the capillary electrophoresis method, sample treatment procedure and sieving medium composition for enhanced shortmers separation of mRNA by CGE analysis. An RNA ladder with 100-1000 nucleotides and EPO mRNA with 900 nucleotides were used as model compounds. The effect of capillary dimensions and separation temperature on the resolution of the RNA peaks was established through comparative experiments. Sample treatment processes were evaluated to achieve an optimal conformation of the mRNA for CGE analysis. By heating the mRNA sample for 15 min at 80°C all multimers were seemingly eradicated. Moreover, it was found that addition of 4 M of urea to mRNA sample before heating resulted in improved peak shape. A sieving medium consisting of a mix of the two polymers polyvinylpyrrolidine (PVP) and hydroxyethyl cellulose (HEC) proved to have beneficial qualities for separation. The addition of sucrose as viscosity modifier in the sieving medium surprisingly further enhanced the resolution. Moreover, during the project a heavy wash was established which drastically improved repeatability of the analyses through more efficient regeneration of the capillary. ISSN:

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Walton, M. P. "The application of gel-electrophoresis to the study of cereal aphid parasitoids." Thesis, University of Hertfordshire, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.373437.

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Wong, Vicky. "Analysis of Arabidopsis xylans using polysaccharide analysis of carbohydrates using gel electrophoresis." Thesis, University of Cambridge, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.613790.

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Goldman, Johnathan M. "Direct MicroRNA Detection Using Kilobase DNA Nantoags In Rapid Gel- Free Electrophoresis." Research Showcase @ CMU, 2015. http://repository.cmu.edu/dissertations/571.

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The abnormal expression of microRNA (miRNA) within tissues has been linked to the onset of many diseases and cancers. The ability to employ miRNA as biomarkers for disease requires sensitive, high-throughput detection, ideally without expensive indirect enzymatic processing. We present our development of a direct, sensitive sandwich hybridization miRNA detection assay. We achieve hybridization of two probes to the short miRNA targets using high affinity PEG γ- carbon modified peptide nucleic acid amphiphiles (γPNAA). The γPNAA enables hybridization of a second, highly fluorescent DNA probe stained with intercalating dye, termed a nanotag, for sensitive detection of the low abundance miRNAs. Upon hybridization of both probes, an electrophoretic mobility shift is measured via interaction of the hydrocarbon modification of the γPNAA with non-ionic surfactant micelles in a capillary electrophoresis running buffer, a technique known as micelle end-labeled free-solution electrophoresis (ELFSE). We demonstrate multiplexed detection of 6 let-7 miRNAs in 4 minutes, excellent selectivity against G-T wobble base-pair single base mismatches between let-7 miRNAs, and 100pM detection limits using our method. In an effort to increase detection sensitivity closer to those required for trace miRNA concentrations (fM), we have investigated the use of isotachophoretic injections and longer nanotags. Although longer nanotags can accommodate a greater amount of fluorescent dye for enhanced signal, they are a challenge to separate by micelle ELFSE. The longer DNA lengths overcome the additional friction of the transiently attached micelle and return to the inseparable freesolution limit. We find that n-alkyl polyoxyethylene ether surfactant (CiEj) wormlike micelles yield high drag forces on the electrophoresing DNAs, appearing to be ideal for large nanotag separations. However, micelle-micelle interaction at moderate concentrations leads to a background sieving network that reduces ELFSE separation efficiency for long DNAs. We propose new constitutive models that accurately capture the behavior of long DNA lengths in these micelle networks. We demonstrate optimization of surfactant buffers to achieve separation of 1-10 kilobase DNA in 3 minutes and increase DNA sequencing length of reads by refining the degree of micelle-micelle interaction. Finally, we demonstrate micelle ELFSE separations on a microchip format for dramatically reduced separation times.

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Romagnolo, Donato. "Ruminal degradability of subfractions of protein sources as determined by gel electrophoresis." Thesis, Virginia Tech, 1988. http://hdl.handle.net/10919/45176.

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Degradability in the rumen of several protein sources was determined by suspending from 12 to 13 g of feedstuff in dacron bags into the rumen for 0, 2, 6, 12, 24, 36, 48, and 72 h. Rumen cannulated lactating Holstein cows consuming a diet of corn silage, alfalfa, soybean, and high moisture corn were used. Degradability of protein varied from 18.6% for corn gluten meal to 72.3% for soybean meal. Gel electrophoresis was used to monitor rates of degradation in the rumen of fractions of corn gluten (CGM), CORN, cottonseed (CSM), peanut (PM), and soybean meal (SBM) protein fractions. Fractional degradation rates in the rumen were determined from densitometric analysis of stained polypeptides bands on SDS-PAGE gels. Acidic subunits of soybean glycinin were degraded at a faster rate than basic subunits (.144 vs .104 h⁻¹). Rates of degradation of zein in corn and corn gluten meal were .026 and .015 h⁻¹, respectively. Protein degradability estimated by using B subfractional components did not differ from degradability measured using total B fractions. Lag phase associated with dacron bags suspension technique did not change effective degradability. Protein solubility in SDS-PAGE sample buffer was highly correlated (R²=.958) with in situ protein degradability of CORN, CSM, DBG, FM, PM, and SBM. Different rates of degradation of each fraction may directly influence protein and amino acid contribution to the animal.
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Blom, Dirk Jacobus. "Polyacrylamide gradient gel electrophoresis for the diagnosis of dysbetalipoproteinaemia (Type III hyperlipidaemia)." Master's thesis, University of Cape Town, 2001. http://hdl.handle.net/11427/3365.

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Includes bibliographical references.
Accurate phenotypic diagnosis of this disorder has generally relied on the quantitative analysis of centrifugally isolated VLDL. Analysis of VLDL chemical composition is not widely available due to the expense of acquiring ultracentrifuges and the highly-skilled staff necessary to perform the analyses. Genotypic diagnosis is somewhat more widely available, especially testing for the E2/E2 genotype which is the commonest underlying genotype in dysbetalipoproteinaemia. Testing for the rarer autosomal mutations of ApoE is restricted to a small number of centres only. In this study I will review dysbetalipoproteinaemia briefly and then describe and evaluate GGE as a new diagnostic technique for this disorder.

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Hall, Richard Lee. "Physical mapping and field inversion gel electrophoresis of Amsacta moorei entomopoxvirus DNA /." The Ohio State University, 1989. http://rave.ohiolink.edu/etdc/view?acc_num=osu148767311411527.

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Wang, Dayou. "Information extraction from DNA pulsed-field gel electrophoresis patterns and it's application /." free to MU campus, to others for purchase, 2000. http://wwwlib.umi.com/cr/mo/fullcit?p9988709.

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Anduri, Sridevi. "Differential protein expression profiles in normal and intersex male smallmouth bass determined using one- and two-dimensional gel electrophoresis a thesis presented to the faculty of the Graduate School, Tennessee Technological University /." Click to access online, 2009. http://proquest.umi.com/pqdweb?index=0&did=2000377671&SrchMode=1&sid=2&Fmt=6&VInst=PROD&VType=PQD&RQT=309&VName=PQD&TS=1277817194&clientId=28564.

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Eumkeb, Griangsak. "Investigation of the effect of antifolates on Escherichia coli 1810." Thesis, Robert Gordon University, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.265230.

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Warrender, Garry William. "Some structure, property and function relations of polyacrylamide flocculants / Garry William Warrender." Thesis, The University of Sydney, 2005. https://hdl.handle.net/2123/28020.

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In this work, the question of how the structure-property relations of polyacrylamide flocculants relate to the flocculation of natural macromolecules and colloids is posed. The question requires an understanding of the chemical function of the polymers and, subsequently, an understanding of how the polymers are binding to the substrate that is flocculated.

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Pandey, Archana. "Proteome analysis of Pseudomonas putida KT2440 using 2D gel electrophoresis and LC/ESI-Q-TOF mass spectrometry /." Online version of thesis, 2007. https://ritdml.rit.edu/dspace/handle/1850/3848.

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Drinnon, Damon L. J. "Use of pulsed-field gel electrophoresis to genotypically characterize salmonellae grouped by serotype." Thesis, Texas A&M University, 2003. http://hdl.handle.net/1969.1/2185.

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The prevention and control of salmonellae in commercial swine operations are becoming increasingly important. The current approach focuses on identifying sources and/or origins of salmonellae contamination before swine are processed for human consumption. The objective of the current study was to assess strain variability among salmonellae grouped by serotype and to determine common origins of contamination (farm or slaughter plant). Salmonellae were previously collected from swine at slaughter, serotyped by the National Veterinary Services Laboratory and stored at - 70??C. Pulsed-field gel electrophoresis (PFGE) was performed to genotypically characterize serotypic isolates using restriction endonuclease XbaI. Dendrogram comparisons were also used to assess genotypic similarity when multiple genotypes existed. This study found PFGE to be more discriminatory than serotyping indicating that multiple genotypic strains existed among selected serotypes. On the basis of PFGE results alone, origins of contamination could not be determined in this study. It is suggested by the author, that origins of contamination could be further defined pending future research, in which in-depth longitudinal studies are included. When used as an adjunct to conventional typing methods, PFGE may prove to be a substantial subtyping system in epidemiologic investigations to identify point-of-entry contaminants to the food chain.

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Medeiros, Diane. "Epidemiological typing of Campylobacter clinical and food isolates using pulsed-field gel electrophoresis." Thesis, University of Ottawa (Canada), 2002. http://hdl.handle.net/10393/6279.

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Campylobacteriosis is the most frequently reported type of acute bacterial gastroenteritis in Canada. In 2000 alone, 11,846 campylobacteriosis cases were reported in Canada. The majority of these cases are sporadic, and their causes remain unknown. An attempt was made to gain a better understanding of the epidemiology of campylobacteriosis, both by identifying foods and environments that harbor Campylobacter spp., and by characterizing clinical, food and environmental isolates using pulsed-field gel electrophoresis (PFGE). Spot maps were also used to determine the geographical relationship of these campylobacteriosis cases. A variety of raw and ready-to-eat foods were tested for the presence of Campylobacter spp. From the 300 samples analyzed, Campylobacter spp. were detected in four samples, one raw beef liver sample and three raw chicken samples. An isolation rate of 9.7% was observed among the raw chicken samples tested, a significantly-reduced percentage, as compared to a 1981 Canadian survey. The prevalence of Campylobacter spp. in a poultry foodservice operation in Ottawa, was also determined from March to August 2001. No Campylobacter spp. were detected in the 125 samples tested. Campylobacter clinical and food isolates were characterized using PFGE with two restriction enzymes, SmaI and KpnI. KpnI produced more complex banding patterns than SmaI, and proved to be more discriminatory. Among the 154 isolates assigned to clusters by SmaI, only 42% gave concordant results with KpnI. In contrast, among the 53 isolates assigned to 23 clusters by KpnI, 87% gave concordant results with SmaI. Five of the 20 concordant clusters represented isolates obtained from the same person, suggesting that some of these individuals may have become re-infected. Spot map analysis revealed a significant clustering of campylobacteriosis cases in the former city of Ottawa, most of which, did not belong to the same postal code. In contrast, very few cases were observed in outlying regions; however, most of these cases belonged to the same postal code, suggesting the possible presence of local outbreaks.

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Sean, David. "On the mobility of partially denatured DNA in gel electrophoresis: a theoretical investigation." Thesis, University of Ottawa (Canada), 2010. http://hdl.handle.net/10393/28742.

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There are technologies which exploit a rapid reduction of the gel electrophoretic mobility of DNA arising from partial denaturation. The underlying phenomenon behind these experiments---the mechanisms which reduce the mobility---are not very well understood. Such is the purpose of my thesis. The first chapter provides a brief introduction to the field of polymer physics. The subjects covered are carefully chosen to directly relate to the forthcoming research. There is a published semi-empirical formula used to model the rapid decrease of mobility which is largely considered to be consistent with experimental data. The second chapter of this thesis demonstrates that there is a fundamental confusion in the literature regarding the fitting parameter Lr, in the said formula. By going back to the original derivation, a physical interpretation can be given to L r. This interpretation yields theoretical values which are consistent with what has been published. However, we find that an underlying assumption---that the effect of the denaturation does not depend on its position along the DNA fragment---may systematically overestimate experimental observations of Lr. To measure the impact of this assumption, a simulation model of DNA is presented. The article presented in the third chapter reveals that indeed, the position of the denatured region affects the migration of the DNA fragment. A refined version of the formula which takes these factors into account is proposed. The simulations also reveal that, for certain fields, an unexpected conformation completely dominates during migration of the fragment. This surprising result: a squid-like conformation, is explored in chapter four.

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Litt, Lloyd C. "An investigation of electrophoresis gel silver staining using large area sample inclusive polymerization /." Online version of thesis, 1989. http://hdl.handle.net/1850/11463.

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37

Lang, Alastair Michael. "Developing tissue proteomics : differential in gel electrophoresis in biomarker discovery and proteomic degradation." Thesis, University of Glasgow, 2013. http://theses.gla.ac.uk/4642/.

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The field of proteomics and functional genomics has developed steadily since the completion of the human genome project. The wealth of genomic information and the pace at which it was compiled was astounding. Proteomics, despite considerable effort, on the other hand has not seen quite the same pace of development. The progress being considerably hindered by the lack of an amplification process and the relative complexity of the proteome in comparison to the genome. These intrinsic difficulties have led to the sensitivity of proteomic techniques being pushed closer to physical limits. There is therefore a further need to re-evaluated techniques such as sample preparation and integrity, analytical methods and collaborative strategies to maximise the effectiveness and quality of data collected. The importance of tissue in scientific and clinical research is unequivocal. However, tissue is difficult to collect, store and work with due to issues with proteomic degradation and storage. Good lab practices can minimise the effect of degradation but degradation of proteins can be rapid. Strategies to minimise degradation include freezing, formalin fixing and microwave treatment which all have their relative advantages and disadvantages. The importance of sample preparation as being the top of the workflow is often acknowledged but improvements are not well described in the literature. The main aim of this thesis is to present investigative studies into the mitigation of some of the limitations in tissue sample degradation, analytical approaches in differential in gel electrophoresis and accessing DiGE spot and tissue profile data. Presented is the evaluation of the effectiveness of rapid and controlled heating of intact tissue to inactivate native enzymatic activity and to aid in the cessation of proteomic degradation. A multifaceted analytical approach of differential in Gel electrophoresis spot data is assessed, giving proteomic profiles of mouse brain tissue. Preliminary data is presented showing that the process of heat-treatment has had a predominantly beneficial effect on mouse brain tissue, with a higher percentage of spots stabilised in heat-treated samples compared to snap-frozen samples. However, stabilisation did occur in snap-frozen samples for different protein spot so the appropriateness of using heat-treatment is as yet not fully determined and requires further analysis. In addition, the variation in tissue profiles of WKY, SP.WKYGla.2a and SHRSP rat model for hypertension is investigated with the future prospect of providing that vital connection between genomic and proteomic data and link phenotype and genotype preliminary investigated. A number of putative markers were identified and quantified using DiGE analysis. In order for these markers to be accepted as biomarkers, more downstream validation is required, however this study provides a good spring board as a proof of concept in using DiGE as an global putative biomarker discovery platform.

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Lam, Ching-to, and 林正道. "Molecular characterization of clostridium difficile isolates by capillary gel electrophoresis-based PCR ribotyping." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2012. http://hub.hku.hk/bib/B48333992.

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Backgroud: Clostridium difficile infection (CDI) is a global concern since the emergence of the hyper-virulent strain NAP1/BI/027. The incidence of CDI has been increasing and active surveillance of Clostridium difficile epidemiology is of paramount importance. In studying the epidemiology of CDI, PCR ribotyping is a very useful method in finding the genetic relatedness of outbreak and epidemic strains. Characterisation of Clostridium difficile isolates can help surveillance and active monitoring, hence reducing the chance of potential outbreaks. Aim: The aims of this study is to investigate the epidemiology of Clostridium difficile in Hong Kong in the year 2010, and that of a Clostridium difficile outbreak in 2011 by using capillary gel electrophoresis-based PCR ribotyping. Results: Among the 307 toxigenic isolates, 139 isolates (45.3%) were characterized as existing ribotypes. Ribotype 012 was the predominant ribotype (17.3%), followed by ribotype 002/0 (15.6%). A total of 144 isolates (46.9%) were characterized as new ribotypes. The remaining 24 isolates (7.8%) were characterized as of high resemblance of existing ribotypes. A total of 8/12 isolates (66.7%) of the outbreak strains were found to be ribotype 002/0. Conclusion: The predominant strains of Clostridium difficile in 2010 was ribotype 012. The 2011 outbreak in Kowloon Hospital was an outbreak of Clostridium difficile ribotype 002/0. Capillary gel electrophoresis-based PCR ribotyping was showed to be a useful tool in investigation of Clostridium difficile outbreaks and study of Clostridium difficile epidemiology.
published_or_final_version
Microbiology
Master
Master of Medical Sciences

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39

Cadiou, Helene. "Evaluation of denaturing gradient gel electrophoresis (DGGE) as a method of mutation detection." Thesis, University College London (University of London), 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.418093.

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40

Le, Riche Mia. "Lipoprotein X : biochemical predictors and detection by non-denaturing polyacrylamide gradient gel electrophoresis." Thesis, Stellenbosch : Stellenbosch University, 2007. http://hdl.handle.net/10019.1/18218.

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Assignment (MMed)--University of Stellenbosch, 2007.
ENGLISH ABSTRACT: Lipoprotein X (LpX) is an abnormal cholesterol-containing particle that may be present in the serum of subjects with cholestasis, lecithin:cholesterol acyltransferase (LCAT) deficiency and parenteral nutrition. The biochemistry, metabolism, clinical significance and laboratory analysis of LpX is discussed in this study. This laboratory-based project investigated icteric samples received at the Chemical Pathology laboratory, Tygerberg Hospital, for serum predictors of LpX and the use of a modified non-denaturing polyacrylamide gradient gel electrophoresis system in the detection of LpX. The study showed that the non-denaturing polyacrylamide gradient gel electrophoresis system (2-8%) is a useful test in demonstrating LpX in icteric plasma and has potential for a screening test in LCAT deficiency. Serum concentration of conjugated bilirubin, alkaline phosphatase, gamma glutamyltransferase, free cholesterol, phospholipid, free cholesterol: total cholesterol ratio and conjugated bilirubin: total bilirubin ratio are all good predictors of LpX. The ratio of free cholesterol to total cholesterol (FC/TC > 0.6) was the best predictor of LpX. In the setting of obstructive liver disease LpX is seen in 66% of patients if total cholesterol is > 7.5 mmol/L.
AFRIKAANSE OPSOMMING: Lipoproteien X (LpX) is ‘n abnormale cholesterol-bevattende partikel wat teenwoordig mag wees in die serum van persone met cholestase, lesitien:cholesterol asieltransferase (LCAT) gebrek en parenterale voeding. Die biochemie, metabolisme, kliniese belang en laboratorium analise van LpX word bespreek in hierdie werkstuk. Hierdie laboratorium-gebaseerde projek het geelsugtige monsters ondersoek wat ontvang is by die Chemiese Patologie laboratorium, Tygerberg Hospitaal, vir serum voorspellers van LpX en die gebruik van ‘n gemodifiseerde nie-denaturerende polie-akrielamied gradiënt gel elektroforese sisteem in die demonstrasie van LpX. Die bevindinge was dat die nie-denaturerende polie-akrielamied gradient gel elektroforese sisteem (2-8%) is ‘n nuttige toets om LpX te demonstreer in geelsugtige plasma en het potensiaal as ‘n siftingstoets in LCAT gebrek. Serum konsentrasie van gekonjugeerde bilirubien, alkaliese fosfatase, gamma glutamieltransferase, vry cholesterol, fosfolipied, vry cholesterol:totale cholesterol verhouding en gekonjugeerde bilirubien:totale bilirubien verhouding is alles goeie voorspellers van LpX. Die verhouding van vry cholesterol tot totale cholesterol (VC/TC > 0.6) was die beste voorspeller van LpX. In gevalle van obstruktiewe lewersiekte word LpX gesien in 66% van pasiente as die totale cholesterol meer as 7.5 mmol/l is.

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Tomlinson, Kerry. "Speciation of metals in proteins using gel electrophoresis with laser ablation-ICP-MS." Thesis, University of Sheffield, 2003. http://etheses.whiterose.ac.uk/14729/.

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A study into the applicability of laser ablation inductively coupled plasma-mass spectrometry (LA ICP-MS) coupled with native polyacrylamide gel electrophoresis (PAGE) to metal speciation in proteins is described in this thesis. Chapter one of the thesis outlines the various roles played by metal ions in clinical samples. Health risks can arise from anthropogenic metal enrichment and metallodrug therapy is identified as a key source. The use of platinum and gold metallodrugs is discussed in detail. Speciation of metals in clinical samples is examined, concentrating on some of the techniques currently employed and their limitations. Particular attention is paid to the techniques employed in the study. The aim of the study is outlined - to develop an alternative speciation strategy for the study of metals in proteins. The reagents and instrumentation used are described in chapter two along with all experimental procedures. Chapter three describes the method development and determination of the analytical performance of the technique. The technique is used to study platinum speciation of protein samples enriched in vitro, the metal distribution profiles obtained are shown. The technique is next applied to the analysis of in vivo samples obtained from platinum therapy patients and a control source. Chapter four outlines the application of the technique to analysis of gold enriched samples both in vitro and in vivo from chrysotherapy patients. The gold distribution profiles obtained are shown and discussed. Chapter five looks at multi-element distribution and interactions occurring in serum. Elements naturally occurring at trace levels in serum, such as Cu and Fe, are studied in vivo. Other, lower level metals are studied following in vitro enrichment. Chapter six concludes the study. It discusses how the technique was found to be applicable for metal speciation of clinical samples by meeting the criteria laid out at the start of the study. Recommendations for future work are also outlined.

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Yao, Mingyi. "Proteomic studies of Pseudomonas putida KT2440 in carcinogenicity screening via 2-dimensional gel electrophoresis /." Online version of thesis, 2005. https://ritdml.rit.edu/dspace/handle/1850/1118.

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Toledo, Érica Gil Hernandes de. "Perfil eletroforético de proteínas séricas e urinárias de cães normais e de portadores de insuficiência renal crônica /." Jaboticabal : [s.n.], 2001. http://hdl.handle.net/11449/95978.

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Orientadora: Marileda Bonafim Carvalho
Banca: Julieta Rodini Engracia Moraes
Banca: Aguemi Kohayagawa
Resumo: Para este estudo foram avaliados 20 cães adultos sadios (10 machos e 10 fêmeas) e 24 cães adultos (16 machos e 8 fêmeas) com insuficiência renal crônica (IRC), com o objetivo de estudar a proteinúria. Foram empregadas técnicas para determinação quantitativa de proteína da urina e para a separação de suas frações por meio de SDS-PAGE. As quantidades de proteínas presentes na urina de cães sadios foram pequenas, mas o estudo dos perfis eletroforéticos mostrou haver diferenças entre machos e fêmeas. Apesar da ampla variação das concentrações de proteínas entre os animais, o grupo dos portadores de IRC apresentou proteinúria intensa. Os resultados dos portadores de IRC foram agrupados de acordo com o sexo ou em função do tipo de lesão renal predominante, para comparação dos eletroforetogramas. As análises revelaram que os perfis eletroforéticos seguem um padrão para os machos e outro pra as fêmeas. Ainda, o padrão o perfil eletroforético observado quando há predomínio de lesões glomerulares difere do observado quando há predomínio de lesões túbulo-intersticiais.
Abstract: This trial was conducted in 20 adult health dogs (10 male and 10 female) and in 24 adult dogs (16 male and 8 female) in chronic renal failure (CRF), in order to study urinary protein loss. Determination of total urinary proteins and their separation with SDS-PAGE were employed. The results show that CRF dogs had higher proteinuria when compared to controls and the amount of detected protein had large difference among them. In the CRF there are different patterns of protein bands occurrence and distribution according to the patient sex and predominant renal lesion. The difference between protein bands occurrence of male and female already exists in normal dogs.
Mestre

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44

Ethell, Douglas Wayne. "Analysis of developing chick Gallus domesticus spinal cord proteins using two dimensional gel electrophoresis." Thesis, University of British Columbia, 1990. http://hdl.handle.net/2429/29834.

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Several recent experiments on developing chick spinal cord have established a time window when the developing spinal cord changes from a permissive to a restrictive environment for regeneration. This time window occurs during embryonic days 13-14 (E13-E14) of chick development. Recent experiments in adult rat, have found two proteins that actively inhibit axonal regeneration. This study has sought possible inhibitory proteins, in chicks, correlating to this temporal change. Proteins continuously present after this change (E14-E20) but not before (E11) were identified. Two-dimensional gel electrophoresis was used for separatation of the proteins. Seven protein spots of interest demonstrated this correlative late-expressing neural protein (LNP) profile. Although the functions of these proteins could not be ascertained in this study, further investigation is warranted.
Science, Faculty of
Zoology, Department of
Graduate

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Lannon, Christopher L. "Single-cell gel electrophoresis, a potential tool for assessment of in vitro chemotherapeutic sensitivity." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/tape17/PQDD_0001/MQ36486.pdf.

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Rajagopal, Meena Uma. "METHODS DEVELOPMENT AND APPLICATION OF TWO-DIMENSIONAL GEL ELECTROPHORESIS AND MASS SPECTROMETRY IN PROTEOMICS." UKnowledge, 2006. http://uknowledge.uky.edu/gradschool_diss/292.

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The development of a highly sensitive ruthenium-based fluorescent staining solution isdescribed in this dissertation. The in-house synthesized ruthenium complex (RuMS)containing both sulfonated and non-sulfonated ligand has detection limit of 1 ng ofprotein that is better than colloidal coomassie, silver and ruthenium complex containingall sulfonated ligands (RuBPS). RuMS stain has 100-fold dynamic range and does notinterfere with subsequent mass spectral identification of proteins. The capability of inhousesynthesis of the staining solution makes it a viable cost-effective alternative to theexpensive commercially available fluorescent stain, Sypro Ruby. The low detection limit,broad linear dynamic range and compatibility with mass spectrometry, make thedevelopment of this stain a worthwhile pursuit. The staining solution was utilized insubsequent applications of two-dimensional gel electrophoresis (2-DE) technology.Proteomics methodology utilizing 2-DE and mass spectrometry was applied toinvestigate the effect of malathion on the proteome of human neuroblastoma cells.Results indicated that out of 122 proteins that were identified from the neuroblastomaproteome, sixteen proteins were down-regulated while five proteins were significantlyup-regulated after treatment with malathion. Significant down-regulation of calciummodulators like calmodulin and calgizarrin and other key chaperones makes themalathion-treated cells highly prone to oxidative stress. With increased awareness inpesticide related adverse effects, identification of altered proteins in malathion-treatedhuman neuroblastoma cells is a critical finding.Proteomics is a major area of research in the identification of biomarkers for diseases. Anovel immunoprecipitation method developed in this work allowed for successfulisolation and identification of albumin-interactome in cerebrospinal fluid (CSF) that isusually under-represented in standard CSF analysis using 2-DE. A key finding is thedifferential expression of various isoforms of proteins in CSF albumin-interactome fromAlzheimer's disease (AD) subjects. The data implicate the acidic isoform ofprostaglandin D2 synthase (PGDS2) as a potential biomarker for AD. An understandingof the differential expression of these protein isoforms in AD will provide insight into theetiology of the disease and this can have far-reaching implication on drug developmentleading to the cure or even preventation of the disease.

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Huysamen, Cristal. "Diversity in the applications of the single cell gel electrophoresis (comet) assay / Cristal Huysamen." Thesis, North-West University, 2005. http://hdl.handle.net/10394/586.

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The development of the single cell gel electrophoresis assay (Comet assay) as a powerful method for measuring DNA strand breakage and repair, has lead to a broader understanding of the impact of certain internal and external factors on DNA damage. This study describes the establishment of the Comet assay in our laboratory and its application in a diversity of studies. These studies include the monitoring of the effect of exercise on DNA damage and repair with the purpose of establishing the optimal exercise program for middle aged men; in the analysis of DNA repair in a breast cancer patient after chemotherapy, we demonstrated that to get a really informative picture of baseline DNA damage, it is necessary to invoke some type of insult to the DNA; we also studied the antioxidant activity of plant extract and finally an industrial application of the Comet assay where we studied DNA damage and repair in the lymphocytes of people exposed to a variety of chemicals. The Comet assay was made even more informative by incorporating additional steps by digesting the DNA on the microscope slides with enzymes that recognize particular kinds of damage to the nucleic acid (formamidopyrimidine DNA glycosylase (Fpg) and Endonuclease Ill (Endo Ill). From these applications we can safely conclude that we have established the Comet assay as reproducible assay with proven diversity in its applications. The most significant contribution of this study is the quite novel modification of the Comet assay to monitor the methylation status of DNA. In this we applied the restriction endonuclease enzymes, Hpa ll and Msp I, which have different sensitivities to methylation of their common target site. Changes in the methylation levels of DNA after treatment of animals and isolated cells with chemicals with known effects on DNA, were convincingly demonstrated and indications of cell specificity in DNA methylation were observed. We believe that this modification opens a new field of applications for the Comet assay.
Thesis (M.Sc. (Biochemistry))--North-West University, Potchefstroom Campus, 2005.

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Wilson, Anthony H. "The use of pulsed field gel electrophoresis in the genetic analysis of oilseed rape." Thesis, University of Wolverhampton, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.306957.

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Tevis, Denise Susanne. "Heterocyclic Diamidines Induce Sequence Dependent Topological Changes in DNA; A Study Using Gel Electrophoresis." unrestricted, 2009. http://etd.gsu.edu/theses/available/etd-04162009-154105/.

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Thesis (M.S.)--Georgia State University, 2009.
Title from file title page. W. David Wilson, committee chair; Stewart A. Allison, Kathryn B. Grant, committee members. College of Arts and Sciences.Description based on contents viewed July 22, 2009. Includes bibliographical references (p. 85-87).

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Milkevitch, Matthew. "Mixed-Metal Ruthenium-Platinum Polyazine Supermolecules: Synthesis, Characterization and Exploration of DNA Binding." Diss., Virginia Tech, 2000. http://hdl.handle.net/10919/28142.

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The goal of this research was to design, prepare and study a new class of supermolecules coupling ruthenium and platinum, which would display covalent binding to DNA. Drawing upon the well-established efficacy of cis-diamminedichloroplatinum(II) (cisplatin) and the DNA-binding properties of select ruthenium polyazine complexes, the approach was to bind the cis-PtIICl2 active site of cisplatin to ruthenium light absorbers using the dpq and dpb bridging ligands (where dpq = 2,3-bis(2-pyridyl)quinoxaline, dpb = 2,3-bis(2-pyridyl) benzoquinoxaline). These complexes are potentially bifunctional, capable of DNA intercalation through the bridging ligand and covalent binding to DNA through the cis-PtCl2 site. Synthetic methods were developed to prepare the mixed-metal, bimetallic complexes [(bpy)2Ru(BL)PtCl2](CF3SO3)2 and [(phen)2Ru(BL)PtCl2](CF3SO3)2 (where bpy = 2,2Â¢-bipyridine, phen = 1,10-phenanthroline) in high purity and good overall yields. The DNA-binding ability of these complexes was probed by reaction with linearized plasmid DNA and subsequent analysis by native and denaturing gel electrophoresis. The known DNA binders, cisplatin and trans-{[PtCl(NH3)2]2(m-H2N(CH2)6NH2)}(NO3)2 (1,1/t,t), were examined under equivalent conditions and used as positive controls. Native gel electrophoresis was used to show that these complexes strongly bind DNA, retarding the migration of DNA through the gel in a fashion inversely proportional to the ratio of DNA base pairs (bp) to metal complex (mc). Analysis by denaturing gel electrophoresis determined that the Ru-Pt complexes bind to DNA in a fashion similar to cisplatin, forming primarily intrastrand adducts. However, these systems also appear to form interstrand adducts at a 10-fold lower metal concentration than cisplatin. In addition to affecting the migration rate, the bimetallic complexes also significantly reduced the fluorescence of DNA-intercalated ethidium bromide for the Ru-Pt reacted samples at low-DNA bp: mc ratios. This was not observed for the cisplatin and 1,1/t,t treated samples. This observation was quantitated by gel densitometry. Precipitation of the DNA by cisplatin, 1,1/t,t and all four Ru-Pt complexes was determined not to be the cause of reduced ethidium bromide fluorescence intensity. Homogenous solution fluorescence quenching studies have revealed that the Ru-Pt complexes quench the emission of ethidium bromide even in the absence of DNA, whereas cisplatin and 1,1/t,t do not. In order to compare the effects on DNA migration produced by cisplatin, 1,1/t,t and the Ru-Pt complexes, Rf values were calculated. This analysis has revealed that all four Ru-Pt complexes retard DNA migration to approximately the same degree. Calculation of theoretical DNA migration distances, based upon the molecular weight change of DNA caused by metal-complex binding, have revealed that the observed affect on DNA migration cannot be accounted for by an increase in molecular weight alone. This indicates that changes in charge and three-dimensional shape of the DNA upon binding of the Ru-Pt complexes may also contribute.
Ph. D.

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