Relevant bibliographies by topics / Plasmid DNA isolation

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  1. Journal articles
  2. Dissertations / Theses
  3. Books
  4. Book chapters
  5. Conference papers

Academic literature on the topic 'Plasmid DNA isolation'

Author: Grafiati
Published: 4 June 2021
Last updated: 13 February 2022

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Journal articles on the topic "Plasmid DNA isolation"

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Hardianto, Dudi, Alfik Indarto, and Nurtjahjo Dwi Sasongko. "OPTIMASI METODE LISIS ALKALI UNTUK MENINGKATKAN KONSENTRASI PLASMID." Jurnal Bioteknologi & Biosains Indonesia (JBBI) 2, no. 2 (December 5, 2015): 60. http://dx.doi.org/10.29122/jbbi.v2i2.510.

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Plasmids are extra chromosomal molecules of DNA that replicate autonomously and found in prokaryote and eukaryote cells. There are a number of methods that are used to isolate plasmids, such as alkaline lysis, boiling lysis, using cesium chloride, and chromatography. Amongst the disadvantages in plasmid isolation methods are lengthy time especially when handling a large number of samples, high cost, and low purity. Alkaline lysis is the most popular for plasmid isolation because of its simplicity, relatively low cost, and reproducibility. This method can be accomplished in 50 minutes to one hour. In this research, the alkaline lysis method was developed to obtain suitable plasmid for applications in a molecular biology laboratory. The aim of this research was to reduce contaminants and improve yield of plasmid. The result of isolation of pICZA plasmid in Escherichia coli gave the concentration of 3.3 to 3.8 µg/µL with the purity of 1.99.Keywords: Plasmid isolation, pICZ A, Escherichia coli, rapid, alkaline lysis ABSTRAKPlasmid merupakan molekul DNA ekstrakromosomal yang bereplikasi secara mandiri dan ditemukan dalam sel prokariot dan eukariot. Banyak metode yang digunakan untuk isolasi plasmid, seperti: lisis alkali, lisis dengan pemanasan, penggunaan sesium klorida, dan kromatografi. Kelemahan beberapa metode isolasi DNA adalah waktu isolasi yang lama terutama saat isolasi plasmid dalam jumlah banyak, mahal dan kemurniannya yang rendah. Metode lisis alkali merupakan metode yang sangat umum untuk isolasi plasmid karena mudah dilakukan, relatif murah, dan reprodusibilitas. Metode ini dapat dilakukan dalam 50 menit sampai 1 jam. Pada penelitian ini dikembangkan metode lisis alkali untuk memperoleh plasmid yang sesuai untuk penggunaan di laboratorium biologi molekuler. Tujuan dari penelitian ini adalah untuk mengurangi jumlah kontaminan dan meningkatkan konsentrasi plasmid. Hasil isolasi plasmid pICZA dalam Escherichia coli mempunyai konsentrasi antara 3,3 sampai 3,8 µg/µL dan kemurniannya 1,99.Kata Kunci: Isolasi plasmid, pICZ A, Escherichia coli, cepat, lisis alkali
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Kniazeva, Marina. "TRIzol for plasmid DNA isolation." Technical Tips Online 2, no. 1 (January 1997): 76–77. http://dx.doi.org/10.1016/s1366-2120(08)70040-0.

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Srutkowska, S., G. Konopa, and G. Wegrzyn. "A method for isolation of plasmid DNA replication intermediates from unsynchronized bacterial cultures for electron microscopy analysis." Acta Biochimica Polonica 45, no. 1 (March 31, 1998): 233–40. http://dx.doi.org/10.18388/abp.1998_4305.

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Electron microscopy is a powerful technique for analysis of DNA replication intermediates. However, isolation of replicating DNA molecules from living cells is tricky and difficult, especially in the case of small DNA molecules (such as bacterial plasmids) whose initiation of replication is not easily synchronized. Here a relatively simple and rapid method for efficient isolation of replicating plasmid molecules from unsynchronized Escherichia coli cultures is described. The efficiency of this procedure is high enough for electron microscopy analysis of plasmid replication intermediates appearing in living cells in normal growth conditions. Under optimal conditions, using standard procedures of isolation of plasmid DNA, it is possible to achieve a content of only as few as 0.02 percent of replication intermediates in a plasmid DNA sample. The described method allowed us to enrich up to 100-fold the fraction of replication intermediates suitable for microscopic analysis among all plasmid molecules.
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Kusnadi, Mr, Diana Rochintaniawaty, and Diah Kusumawaty. "MENGEMBANGKAN KEMAMPUAN MAHASISWA PENDIDIKAN BIOLOGI DALAM MENGISOLASI PLASMID BAKTERI SEBAGAI PENGAYAAN PRAKTIKUM MIKROBIOLOGI." Jurnal Pengajaran Matematika dan Ilmu Pengetahuan Alam 6, no. 2 (December 8, 2005): 27. http://dx.doi.org/10.18269/jpmipa.v6i2.346.

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This learning cycle focused on developing of Biology education student’s performance in bacteria DNA-plasmid isolation as well as using gel electrophroresis. The aim of the study was to improve student’s performance in operating equipment used in plasmid isolation such as: micropipettes, microsentrifuge, vortex, shaker-incubator, gel electrophoresis, and UV Trans- illuminator camera. Beside that students were also trained to DNA-plasmid isolation base on standard procedure, so students could identify the form of plasmid via the use of gel electrophoresis. This study was conducted in 2 cycle learning processes as an enrichment of microbiology activity, which associated with molecular microbes genetics. Using observation sheet did evaluation. The study revealed that biology education students had good performance in isolating the plasmid and all group were succeed to isolate the plasmid. Beside that there were increased in knowledge ability especially associated with molecular microbes genetics subject, which achievment gain average was 18,3% (Class A) and 35,4% (class B). The DNA isolation activity in microbiology learning shared high contribution as well as increasing of knowledge achievment and performance ability.Keyword: DNA-plasmid, gel electrophoresis, isolation, learning cycle
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Williams, Laura E., Chris Detter, Kerrie Barry, Alla Lapidus, and Anne O. Summers. "Facile Recovery of Individual High-Molecular-Weight, Low-Copy-Number Natural Plasmids for Genomic Sequencing." Applied and Environmental Microbiology 72, no. 7 (July 2006): 4899–906. http://dx.doi.org/10.1128/aem.00354-06.

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ABSTRACT Sequencing of the large (>50 kb), low-copy-number (<5 per cell) plasmids that mediate horizontal gene transfer has been hindered by the difficulty and expense of isolating DNA from individual plasmids of this class. We report here that a kit method previously devised for purification of bacterial artificial chromosomes (BACs) can be adapted for effective preparation of individual plasmids up to 220 kb from wild gram-negative and gram-positive bacteria. Individual plasmid DNA recovered from less than 10 ml of Escherichia coli, Staphylococcus, and Corynebacterium cultures was of sufficient quantity and quality for construction of high-coverage libraries, as shown by sequencing five native plasmids ranging in size from 30 kb to 94 kb. We also report recommendations for vector screening to optimize plasmid sequence assembly, preliminary annotation of novel plasmid genomes, and insights on mobile genetic element biology derived from these sequences. Adaptation of this BAC method for large plasmid isolation removes one major technical hurdle to expanding our knowledge of the natural plasmid gene pool.
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Sobecky, Patricia A., Tracy J. Mincer, Michelle C. Chang, Aresa Toukdarian, and Donald R. Helinski. "Isolation of Broad-Host-Range Replicons from Marine Sediment Bacteria." Applied and Environmental Microbiology 64, no. 8 (August 1, 1998): 2822–30. http://dx.doi.org/10.1128/aem.64.8.2822-2830.1998.

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ABSTRACT Naturally occurring plasmids isolated from heterotrophic bacterial isolates originating from coastal California marine sediments were characterized by analyzing their incompatibility and replication properties. Previously, we reported on the lack of DNA homology between plasmids from the culturable bacterial population of marine sediments and the replicon probes specific for a number of well-characterized incompatibility and replication groups (P. A. Sobecky, T. J. Mincer, M. C. Chang, and D. R. Helinski, Appl. Environ. Microbiol. 63:888–895, 1997). In the present study we isolated 1.8- to 2.3-kb fragments that contain functional replication origins from one relatively large (30-kb) and three small (<10-kb) naturally occurring plasmids present in different marine isolates. 16S rRNA sequence analyses indicated that the four plasmid-bearing marine isolates belonged to the α and γ subclasses of the classProteobacteria. Three of the marine sediment isolates are related to the γ-3 subclass organisms Vibrio splendidusand Vibrio fischeri, while the fourth isolate may be related to Roseobacter litoralis. Sequence analysis of the plasmid replication regions revealed the presence of features common to replication origins of well-characterized plasmids from clinical bacterial isolates, suggesting that there may be similar mechanisms for plasmid replication initiation in the indigenous plasmids of gram-negative marine sediment bacteria. In addition to replication inEscherichia coli DH5α and C2110, the host ranges of the plasmid replicons, designated repSD41, repSD121, repSD164, and repSD172, extended to marine species belonging to the generaAchromobacter, Pseudomonas,Serratia, and Vibrio. While sequence analysis of repSD41 and repSD121 revealed considerable stretches of homology between the two fragments, these regions do not display incompatibility properties against each other. The replication origin repSD41 was detected in 5% of the culturable plasmid-bearing marine sediment bacterial isolates, whereas the replication origins repSD164 and repSD172 were not detected in any plasmid-bearing bacteria other than the parental isolates. Microbial community DNA extracted from samples collected in November 1995 and June 1997 and amplified by PCR yielded positive signals when they were hybridized with probes specific for repSD41 and repSD172 replication sequences. In contrast, replication sequences specific for repSD164 were not detected in the DNA extracted from marine sediment microbial communities.
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Rosenshine, Ilan, and Moshe Mevarech. "Isolation and partial characterization of plasmids found in three Halobacterium volcanii isolates." Canadian Journal of Microbiology 35, no. 1 (January 1, 1989): 92–95. http://dx.doi.org/10.1139/m89-014.

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Three new isolates of Halobacterium volcanii were screened for the presence of plasmids. Each of the different isolates was found to contain one plasmid. These plasmids do not show any homology to each other, nor to the previously isolated plasmid pHV2. Partial restriction maps of these plasmids were determined. One of the plasmids contains chromosomal repetitive sequences as judged by the existence of homologous sequences in the chromosomal DNA of the three isolates. Using the protoplast fusion technique, we showed that at least one of the newly isolated plasmids is compatible with pHV2.Key words: Halobacterium volcanii, archaebacterium, plasmids.
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Asmundson, Roderick V., and William J. Kelly. "Isolation and characterization of plasmid DNA fromRuminococcus." Current Microbiology 16, no. 2 (March 1987): 97–100. http://dx.doi.org/10.1007/bf01588178.

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Erickson, J. R., and M. Johnston. "Direct cloning of yeast genes from an ordered set of lambda clones in Saccharomyces cerevisiae by recombination in vivo." Genetics 134, no. 1 (May 1, 1993): 151–57. http://dx.doi.org/10.1093/genetics/134.1.151.

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Abstract We describe a technique that facilitates the isolation of yeast genes that are difficult to clone. This technique utilizes a plasmid vector that rescues lambda clones as yeast centromere plasmids. The source of these lambda clones is a set of clones whose location in the yeast genome has been determined by L. Riles et al. in 1993. The Escherichia coli-yeast shuttle plasmid carries URA3, ARS4 and CEN6, and contains DNA fragments from the lambda vector that flank the cloned yeast insert. When yeast is cotransformed with linearized plasmid and lambda clone DNA, Ura+ transformants are obtained by a recombination event between the lambda clone and the plasmid vector that generates an autonomously replicating plasmid containing the cloned yeast DNA sequences. Genes whose genetic map positions are known can easily be identified and recovered in this plasmid by testing only those lambda clones that map to the relevant region of the yeast genome for their ability to complement the mutant phenotype. This technique facilitates the isolation of yeast genes that resist cloning either because (1) they are underrepresented in yeast genomic libraries amplified in E. coli, (2) they provide phenotypes that are too marginal to allow selection of the gene by genetic complementation or (3) they provide phenotypes that are laborious to score. We demonstrate the utility of this technique by isolating three genes, GAL83, SSN2 and MAK7, each of which presents one of these problems for cloning.
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Nakashima, Nobutaka, and Tomohiro Tamura. "Isolation and Characterization of a Rolling-Circle-Type Plasmid from Rhodococcus erythropolis and Application of the Plasmid to Multiple-Recombinant-Protein Expression." Applied and Environmental Microbiology 70, no. 9 (September 2004): 5557–68. http://dx.doi.org/10.1128/aem.70.9.5557-5568.2004.

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ABSTRACT We isolated, sequenced, and characterized the cryptic plasmid pRE8424 from Rhodococcus erythropolis DSM8424. Plasmid pRE8424 is a 5,987-bp circular plasmid; it carries six open reading frames and also contains cis-acting elements, specifically a single-stranded origin and a double-stranded origin, which are characteristic of rolling-circle-replication plasmids. Experiments with pRE8424 derivatives carrying a mutated single-stranded origin sequence showed that single-stranded DNA intermediates accumulated in the cells because of inefficient conversion from single-stranded DNA to double-stranded DNA. This result indicates that pRE8424 belongs to the pIJ101/pJV1 family of rolling-circle-replication plasmids. Expression vectors that are functional in several Rhodococcus species were constructed by use of the replication origin from pRE8424. We previously reported a cryptic plasmid, pRE2895, from R. erythropolis, which may replicate by a θ-type mechanism, like ColE2 plasmids. The new expression vectors originating from pRE8424 were compatible with those derived from pRE2895. Coexpression experiments with these compatible expression vectors indicated that the plasmids are suitable for the simultaneous expression of multiple recombinant proteins.
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Dissertations / Theses on the topic "Plasmid DNA isolation"

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Nam, Kiebang. "Rapid Isolation and Purification of Plasmid DNA Using High Performance Liquid Chromatography." Thesis, North Texas State University, 1987. https://digital.library.unt.edu/ark:/67531/metadc504136/.

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High Performance Liquid Chromatography (HPLC) has been employed as an analytical tool for the purification and separation of nucleic acids. A Nucleogen DEAE 4000-10 weak anion exchange column, prepacked with modified silica gels, was used to purify and separate a number of Escherichia coli plasmids. Plasmid DNAs were extracted by the alkaline lysis method. The cleared lysate was injected directly onto the Nucleogen column, and the peaks were collected, desalted and analysed by gel electrophoresis. On the chromatogram, the pBR322 formed a distinctive peak at 27 minutes and partial separation was made for the E. coli V517 plasmids. Plasmid pBR322 showed a clear band without any detectable contamination on agarose gel. This purified plasmid DNA is biologically active for enzymatic reaction commonly used in genetic engineering techniques.
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Chlopková, Barbora. "Optimalizace izolace plazmidové DNA pomocí magnetických částic." Master's thesis, Vysoké učení technické v Brně. Fakulta chemická, 2019. http://www.nusl.cz/ntk/nusl-401929.

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The theoretical part summarizes information on the isolation and purification of plasmid DNA and nucleic acids as. Plasmid DNA is often used in gene engineering as a vector for the transfer of a particular gene. Its insulation and transportation in sufficient quality is crucial for other processes associated with it. Isolation and survival of pDNA using magnetic carriers of different concentrations of PEG 8000 in combination with 1M NaCl was investigated in experimental parts. Furthermore, the isolation of pDNA using commercial kits was examined.
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Chen, Jin-Shu, and 陳錦書. "Isolation and characterization of plasmid DNA from a virulent strain of Rhizoctonia solani Kuhn AG-4." Thesis, 1991. http://ndltd.ncl.edu.tw/handle/62581709467358655512.

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Books on the topic "Plasmid DNA isolation"

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Ranger, Nicole J. The isolation of plasmid DNA from a native isolate of Thiobacillus ferrooxidans. Sudbury, Ont: Laurentian University, 1991.

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Manocha, Marcus M. S. Isolation and characterization of genomic DNA sequences that enhance the stability of plasmid DNA in mammalian cells. St. Catharines, Ont: Brock University, Centre for Biotechnology, 2005.

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Book chapters on the topic "Plasmid DNA isolation"

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Boffey, Stephen A. "Isolation of Plasmid DNA." In Springer Protocols Handbooks, 7–15. Totowa, NJ: Humana Press, 1986. http://dx.doi.org/10.1007/978-1-60327-405-0_2.

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Surzycki, Stefan. "Isolation of Plasmid DNA." In Basic Techniques in Molecular Biology, 101–18. Berlin, Heidelberg: Springer Berlin Heidelberg, 2000. http://dx.doi.org/10.1007/978-3-642-56968-5_5.

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Tagg, Kaitlin A., Carola Venturini, Muhammad Kamruzzaman, Andrew N. Ginn, and Sally R. Partridge. "Plasmid DNA Isolation and Visualization: Isolation and Characterization of Plasmids from Clinical Samples." In Horizontal Gene Transfer, 3–20. New York, NY: Springer US, 2019. http://dx.doi.org/10.1007/978-1-4939-9877-7_1.

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Esteban-Torres, Maria, Lorena Ruiz, Rocio Sanchez-Gallardo, and Douwe van Sinderen. "Isolation of Chromosomal and Plasmid DNA from Bifidobacteria." In Methods in Molecular Biology, 21–29. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1274-3_3.

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Lindgren, Jill K. "Isolation of Chromosomal and Plasmid DNA from Staphylococcus epidermidis." In Methods in Molecular Biology, 113–18. Totowa, NJ: Humana Press, 2013. http://dx.doi.org/10.1007/978-1-62703-736-5_9.

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Stray, James, and Bernhard Zimmermann. "Isolation of Cell-Free DNA from Maternal Plasma." In Prenatal Diagnosis, 309–23. New York, NY: Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-8889-1_21.

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Fleischhacker, Michael, Bernd Schmidt, Sabine Weickmann, Debora M. I. Fersching, Gloria S. Leszinski, Barbara Siegele, Oliver J. Stoetzer, and Stefan Holdenrieder. "Comparison of Nucleosomes and Quantitative PCR Using Diverse DNA Isolation Methods." In Circulating Nucleic Acids in Plasma and Serum, 269–73. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-90-481-9382-0_36.

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Marsavela, Gabriela, Anna Reid, Elin S. Gray, and Leslie Calapre. "Isolation and Quantification of Plasma Circulating Tumor DNA from Melanoma Patients." In Methods in Molecular Biology, 247–63. New York, NY: Springer US, 2021. http://dx.doi.org/10.1007/978-1-0716-1205-7_19.

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Huang, Dorothy J., Susanne Mergenthaler-Gatfield, Sinuhe Hahn, Wolfgang Holzgreve, and Xiao Yan Zhong. "Isolation of Cell-Free DNA from Maternal Plasma Using Manual and Automated Systems." In Prenatal Diagnosis, 203–8. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-066-9_15.

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Krishnan, Rajaram, and Michael J. Heller. "Rapid Isolation and Detection of Cell Free Circulating DNA and Other Disease Biomarkers Directly from Whole Blood." In Circulating Nucleic Acids in Plasma and Serum, 247–57. Dordrecht: Springer Netherlands, 2010. http://dx.doi.org/10.1007/978-90-481-9382-0_34.

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Conference papers on the topic "Plasmid DNA isolation"

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Subhi, Hanan, Fatimah Abdel, and Ihsan Raheem. "Isolation of plasmid DNA from Antibiotics Resistant Staphylococcus aureus." In 2018 International Conference on Pure and Applied Science. Koya University, 2018. http://dx.doi.org/10.14500/icpas2018.mim105.

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Ploos van Amstel, J. K., A. L. van der Zanden, E. Bakker, P. H. Reitsma, and R. M. Bertina. "INDEPENDENT ISOLATION OF HUMAN PROTEIN S cDNA AND THE ASSIGNMENT OF THE GENE TO CHROMOSOME 3." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644638.

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Protein S is a vitamin K-dependent glycoprotein, that serves as a cofactor of activated protein C. A hereditary deficiency in protein S is associated with an increased risk for the development of venous thrombosis. The deficiency is inherited as an autosomal dominant trait. We isolated a cDNA coding for protein S and assigned its gene to chromosome 3.A human liver cDNA library in phage xgtll (complexity 1.2 × 106 , D. Stafford, Chapel Hill) was screened by using immuno-purifiedpolyclonal anti-protein S IgG as a probe. Approximately 1.5 x 10 recombinants of the amplified library were screened. Out of eighteen positive clones one clone was found, after nucleotide sequence analysis, to code for a peptide with a high degree of homology with the carboxy terminal region of the already published bovine protein S. This clone pSP84 (450 bp) was used as a probe to screen a human liver cDNA library in plasmid pUC9. From this library we isolated several positive clones. Clone pSUL5 contained the largest insert (2200 base pairs). Dideoxy sequencing revealed that it codes for 330 amino acids of the carboxy terminal part of protein S. Furthermore, it contained a 1200 base pairs 3' untranslated region. The predicted amino acid sequence did not differ from the published sequence of human protein S, although at the nucleotide level some differences could be detected.Clone pSUL5 was used to localize the protein S gene to its chromosome. The assignment was done by hybridization to Pst I digested DNA from human-hamster c.q. human-mouse somatic cell hybrids. In this way we got strong indication that the protein S gene is located on chromosome 3.
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Ichinose, A., R. E. Bottenus, K. R. Loeb, and E. W. Davie. "ISOLATION AND CHARACTERIZATION OF THE GENES FOR THE a AND b SUBUNITS OF HUMAN COAGULATION FACTOR XIII." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644652.

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Factor XIII (plasma transglutaminase, fibrin stabilizing factor) is a plasma protein that plays an important role in the final stages of blood coagulation and fibrinolysis. The molecule occurs in blood as a tetramer (a2b2) consisting of two a. subunits and two b subunits. Recently, we have determined the amino acid sequences for both the a. and b subunits of human factor XIII by a combination of cDNA cloning and amino acid sequence analysis. cDNAs coding for the a (3.8 Kb) and b (2.2 Kb) subunits were used for the screening of human genomic DNA libraries. Among 12 × 106 recombinant phage, ∼30 have been shown to contain the sequences for the a subunit and ∼10 have been shown to contain the gene for the b subunit of factor XIII. The clones coding for the a. subunit span ∼90 Kb and have been characterized by restriction mapping. Southern blotting, and DNA sequencing. Both 5’ and 3’ ends of the genomic clones correspond to the 5’ and 3’portions of the cDNA for the a.subunit of factor XIII. The DNA sequence revealed that the activation peptide released ^thrombin (amino acid residues 137), the first putative Ca2+ binding region (around residue 251), the active Site Cys (amino acid residue 314), and the second putative Ca2+ binding region (around residue 473) are encoded by separate exons. Accordingly, the intervening sequences may separate the a subunit into functional and structural domains. The gene organization for the b subunit will also be presented. (Supported by NIH Grant HL 16919.)
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Sorber, Laure, Karen Zwaenepoel, Vanessa Deschoolmeester, Geert Roeyen, Sofie Goethals, Christian Rolfo, and Patrick Pauwels. "Abstract 486: A comparison of cfDNA isolation kits: isolation and quantification of cell-free DNA in plasma." In Proceedings: AACR 107th Annual Meeting 2016; April 16-20, 2016; New Orleans, LA. American Association for Cancer Research, 2016. http://dx.doi.org/10.1158/1538-7445.am2016-486.

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Dias, Rajen, Lars Skoglund, and Zhiyong Wang. "Laser Milling Methods for Package Failure Analysis." In ISTFA 2002. ASM International, 2002. http://dx.doi.org/10.31399/asm.cp.istfa2002p0109.

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Abstract Current methods used for package level destructive physical analysis (DPA) such as chemical and mechanical decapsulation methods, reactive ion etching (RIE) and diamond saw x-section methods could potentially result in artifacts such as die cracking, delamination or corrosion when used on complex packaging technologies such as multiple thin die stacked packages with combination of flip chip and wire bond interconnections. Many of the shortcoming of these ubiquitous DPA tools are being addressed by a laser milling approach to DPA. The system described in this paper consists of a ultraviolet (UV) laser used for local micromachining or milling to access package internal features and a near infrared(IR) laser used for precise soldering of fine wires to enable testing and fault isolation. Applications of the laser milling tool described in the paper are 1) Delayering of multilayer printed circuit board (PCB) substrates to expose internal metal traces so that they can be tested to fault isolate the failure without loosing electrical functionality of the product. 2) Silicon milling to expose flip chip interconnections. 3) Package cross sectioning and 4) Plastic package decapsulation.
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Uemoto, Yoshio, Akihiko Hirano, and Daisuke Hirasawa. "Fracture Toughness Evaluation of Carbon Steels in Piping and Valve for Reactor Primary System." In ASME 2017 Pressure Vessels and Piping Conference. American Society of Mechanical Engineers, 2017. http://dx.doi.org/10.1115/pvp2017-65579.

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UK very high integrity (VHI) component classification includes design, manufacturing, and inspection requirements that go beyond those established in ASME BPVC Sec. III Subsection NB [1]. One of these requirements is to ensure the component is tolerant of manufacturing defects. This can be demonstrated using a Defect Tolerance Assessment (DTA) based on two parameters fracture mechanics method. The brittle fracture parameter of this assessment requires the analysis of stress occurring in the component against the plane strain fracture toughness, KIC of the material. This work focuses on the practical determination of KIC for materials chosen for a Boiling Water Reactor (BWR) Main Steam Piping (MSP) and Main Steam Isolation Valve (MSIV), which carbon steel seamless pipe SA-106 Grade C and carbon steel casting SA-216 Grade WCB, are respectively. These materials are usually tested by Charpy impact testing specified in [1], but there are not many studies reporting their KIC, and there is not enough information concerning actual piping and valve materials. Thus the authors implemented fracture toughness testing using J-resistance curve according to ASTM E 1820 [2] for test pipe and test casting block simulating actual MS Piping and MSIV, and evaluated KIC(J) to be used in DTA. KIC(J) is evaluated from elastic-plastic fracture toughness, JIC, gained from the J-resistance curve, and equivalent to KIC [3]. KIC(J) corresponds to KJIc in ASTM E 1820. There were some cases, however, in which valid JIC values could not obtained, because of the materials high toughness, test specimen size limitations, and uneven final crack sizes. When valid JIC can’t be obtained, retesting or remanufacturing would significantly affect plant construction schedule. Hence, alternative evaluation methods by which JIC can certainly be obtained are desired. In this study, the authors focused on two types of alternative JIC evaluation methods. The first one is the Stretch Zone Width (SZW) method, in which JIC is calculated from SZW measurements of crack tip plastic blunting on fracture toughness test specimens. The SZW method was well studied in the 1970s, and experimental data showed a clear correlation between JIC values obtained from J-resistance curves and JIC values obtained from SZW measurements [4]. The second method is by correlation of JIC with the energy absorbed during Charpy testing. As represented by Rolf’s study [5], it has been reported that there are correlations between Charpy absorbed energy and KIC for high tensile strength steels. In this study, the validity of the SZW method was first evaluated by comparing its results with JIC obtained from J-resistance curves. Then, the applicability of the JIC values to DTA of actual products was discussed. Finally, by comparing Charpy absorbed energy and KIC(J), the validity and applicability of KIC determination method with Charpy absorbed energy was discussed.
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High, K. A., J. P. Evans, J. L. Ware, D. W. Stafford, and H. R. Roberts. "HEMOPHILIA B IN CANINES IS DUE TO A POST-TRANSCRIPTIONAL DEFECT." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644017.

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Factor IX is a vitamin K dependent plasma proteinsynthesized in the liver; a deficiency of Factor IX results in hemophilia B. An animal model for hemophilia B exists in dogs; affected animals have severe disease, with activity levels of less than 1%. The purpose of the current study is to determine the molecular basis of canine hemophilia. In previous work, we had shown that the Factor IX gene in hemophilic dogs appeared to be at least partly intact; thus, genomic DNA from normal, carrier and hemophilic dogs, when probed with sequences from the 4th, 7th, and 8th exons of the human gene, gave identical patterns on Southern blot. We have now completed the mapping of the hemophilic gene, using probes from the first, second, third and sixth exons, and have shown it to beentirely intact, that is, free of any large deletions or rearrangements, as determined by Southern blotting. In addition, using the guanidinium thiocyanate technique, we h^ve prepared total RNA from normal and -hemophilic dog livers add analyzed these samples by Northern blotting. The results show that the hemophilic dog synthesizes a Factor IX transcript of approximately 3 kilobases, that is, of the same size asthe normal dog. In addition, baaed on signal intensity, the transcript appears to be produced in roughlyequivalent amounts in the normal and hemophilic dogs.We conclude that the defect responsible for canine hemophilia B interferes with the production of the normal Factor IX protein at a post-transcriptional level. Moreover, since the hemophilic dogs produce Factor IX mRNA it should be possible to elucidate the gene defect in the hemophilic animals by preparing normal and hemophilic canine liver cDNA libraries and isolating and characterizing the respective Factor IX cDNAs.
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8

Kaufman, Randal J., Debra D. Pittman, Louise C. Wasley, W. Barry Foster, Godfrey W. Amphlett, and Alan R. Giles. "DIRECTED MUTAGENESIS IN THE STUDY OF THE REQUIREMENTS FOR FACTOR VIII ACTIVITY IN VITRO AND IN VIVO." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644769.

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Factor VIII is a high molecular weight plasma glycoprotein that functions in the blood clotting cascade as the cofactor for factor DCa proteolytic activation of factor X. Factor VIII does not function proteolytically in this reaction hut itself can be proteolytically activated by other coagulation enzymes such as factor Xa and thrombin. In the plasma, factor VIII exists as a 200 kDa amino-terminal fragment in a metal ion stabilized complex with a 76 kDa carboxy-terminal fragment. The isolation of the cENA for human factor VIII provided the deduced primary amino acid sequence of factor VIIT and revealed three distinct structural domains: 1) a triplicated A domain of 330 amino acids which has homology to ceruloplasmin, a plasma copper binding protein, 2) a duplicated C domain of 150 amino acids, and 3) a unique B domain of 980 amino acids. These domains are arranged as shown below. We have previously reported the B domain is dispensible far cofactor activity in vitro (Toole et al. 1986 Proc. Natl. Acad 5939). The in vivo efficacy of factor VIII molecules harboring the B domain deletion was tested by purification of the wildtype and modified forms and infusion into factor VIII deficient, hemophilic, dogs. The wildtype and the deleted forms of recombinant derived factor VIII exhibited very similar survival curves (Tl/2 = 13 hrs) and the cuticle bleeding times suggested that both preparations appeared functionally equivalent. Sepharose 4B chromatography indicated that both factor VIII molecules were capable of binding canine plasma vWF.Further studies have addressed what cleavages are necessary for activation of factor VIII. The position of the thrombin, factor Xa, and activated protein C (AFC) cleavage sites within factor VIII are presented below, site-directed ENA medicated mutagenesis has been performed to modify the arginine at the amino side of each cleavagesite to an soleucine. In all cases this modification resulted in molecules that were resistant to cleavage by thrombin at the modified site. Modification of the thrombin cleavage sites at 336 and 740 and modification of the factor Xa cleavage site at 1721 resulted in no loss of cofactor activity. Modification of the thrombin cleavage site at either 372 or 1689 destroyed oofactor activity. Modification of the thrombin cleavage site at 336 resulted in a factor VIII having an increased activity, possibly due to resistance to inactivation. These results suggest the requirement of cleavage at residues 372 and 1689 for cofactor activity.
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9

Horikoshi, Satoshi. "IN-LIQUID PLASMA USING MICROWAVE POWER FOR APPLICATIONS." In Ampere 2019. Valencia: Universitat Politècnica de València, 2019. http://dx.doi.org/10.4995/ampere2019.2019.9815.

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More than 30 years have passed since Clements, et. al. succeeded in generating plasma in liquid (in-Liquid plasma: LP). Meanwhile, then plasma generation experiments using AC and DC power sources have been performed in electrolyte solutions. On the other hand, in 2000, by Nomura, et. al., they succeeded in generating plasma in aqueous solution by using microwave as a power source. When the microwave is used as a power source, there is a problem that the electrode is deteriorated and melted by the heat of plasma, and there is a problem that the device cannot be used continuously. We solved this problem using a semiconductor (solid-state) microwave generator. In order to investigate the possibility of using this new plasma, we have applied to wastewater treatment (e.g. degradation of 1,4-dioxane, rhodamine B dye and hypochlorous) and gel synthesis (polyvinylpyrrolidone (PVP) gel and silicone hydrogel gel). The photograph of the LP apparatus is illustrated in Figures 1. The MW generator was constructed using an Ampleon M2A-R semiconductor generator (2.45-GHz; maximal power, 1300 W) coupled to an isolator (air cooling device), a power monitor, a three-stub tuner and a short-circuit plunger. Microwaves continuously irradiated the liquid through the tungsten antenna (dia.: 10 mm; length: 200 mm). The tungsten antenna was isolated from the reactor and the waveguide using a ceramic spacer to irradiate MW in the solution. In the application of LP for wastewater treatment, the model wastewater of rhodamine B dye (RhB) were decomposed by LP irradiation, and degradation efficency of LP method was compared with conventional methods (UV photodegradation, NaClO chemical treatment, UV/NaClO chemical/photodegradation and the UV/TiO2 photocatalytic degradation method). The degradationon rate of LP method was remarkably fastest to conventional methods (Figure 2). In the application of LP for gel-synthesis, synthesizing the polymer-gel (PVP-gel and HySi-gel) was tried by the LP method. This feature of the method can significantly reduce (or eliminate) the initiator and crosslinking agent needed for conventional synthesis. Because these chemicals are very toxic, the LP approach is effective in green chemistry. In addition, it will further extend the application of these gels to the medical field.More than 30 years have passed since Clements, et. al. succeeded in generating plasma in liquid (in-Liquid plasma: LP) [1]. Meanwhile, then plasma generation experiments using AC and DC power sources have been performed in electrolyte solutions. On the other hand, in 2000, by Nomura, et. al., [1], they succeeded in generating plasma in aqueous solution by using microwave as a power source. When the microwave is used as a power source, there is a problem that the electrode is deteriorated and melted by the heat of plasma, and there is a problem that the device cannot be used continuously. We solved this problem using a semiconductor (solid-state) microwave generator [2]. In order to investigate the possibility of using this new plasma, we have applied to wastewater treatment (e.g. degradation of 1,4-dioxane, rhodamine B dye and hypochlorous) and gel synthesis (polyvinylpyrrolidone (PVP) gel and silicone hydrogel gel).More than 30 years have passed since Clements, et. al. succeeded in generating plasma in liquid (in-Liquid plasma: LP) [1]. Meanwhile, then plasma generation experiments using AC and DC power sources have been performed in electrolyte solutions. On the other hand, in 2000, by Nomura, et. al., [1], they succeeded in generating plasma in aqueous solution by using microwave as a power source. When the microwave is used as a power source, there is a problem that the electrode is deteriorated and melted by the heat of plasma, and there is a problem that the device cannot be used continuously. We solved this problem using a semiconductor (solid-state) microwave generator [2]. In order to investigate the possibility of using this new plasma, we have applied to wastewater treatment (e.g. degradation of 1,4-dioxane, rhodamine B dye and hypochlorous) and gel synthesis (polyvinylpyrrolidone (PVP) gel and silicone hydrogel gel).
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