Relevant bibliographies by topics / Real-time methylation-specific PCR

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

Academic literature on the topic 'Real-time methylation-specific PCR'

Author: Grafiati
Published: 4 June 2021
Last updated: 12 February 2022

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Journal articles on the topic "Real-time methylation-specific PCR"

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Cottrell, S. E. "A real-time PCR assay for DNA-methylation using methylation-specific blockers." Nucleic Acids Research 32, no. 1 (January 2, 2004): 10e—10. http://dx.doi.org/10.1093/nar/gnh008.

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Pu, Robert T., Zong-Mei Sheng, Claire W. Michael, Michael G. Rhode, Douglas P. Clark, and Timothy J. O'Leary. "Methylation profiling of mesothelioma using real-time methylation-specific PCR: A pilot study." Diagnostic Cytopathology 35, no. 8 (2007): 498–502. http://dx.doi.org/10.1002/dc.20692.

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Bonanno, Cinzia, Erlet Shehi, Daniel Adlerstein, and G. Mike Makrigiorgos. "MS-FLAG, a Novel Real-Time Signal Generation Method for Methylation-Specific PCR." Clinical Chemistry 53, no. 12 (December 1, 2007): 2119–27. http://dx.doi.org/10.1373/clinchem.2007.094011.

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Abstract Background: Aberrant promoter methylation is a major mechanism for silencing tumor suppressor genes in cancer. Detection of hypermethylation is used as a molecular marker for early cancer diagnosis, as a prognostic index, or to define therapeutic targets for reversion of aberrant methylation. We report on a novel signal generation technology for real-time PCR to detect gene promoter methylation. Methods: FLAG (fluorescent amplicon generation) is a homogeneous signal generation technology based on the exceptionally thermostable endonuclease PspGI. FLAG provides real-time signal generation during PCR by PspGI-mediated cleavage of quenched fluorophores at the 5′ end of double-stranded PCR products. Methylation-specific PCR (MSP) applied on bisulfite-treated DNA was adapted to a real-time format (methylation-specific FLAG; MS-FLAG) for quantifying methylation in the promoter of CDKN2A (p16), GATA5, and RASSF1. We validated MS-FLAG on plasmids and genomic DNA with known methylation status and applied it to detection of methylation in a limited number of clinical samples. We also conducted bisulfite sequencing on these samples. Results: Real-time PCR results obtained via MS-FLAG agreed with results obtained via conventional, gel-based MSP. The new technology showed high specificity, sensitivity (2–3 plasmid copies), and selectivity (0.01% of methylated DNA) on control samples. It enabled correct prediction of the methylation status of all 3 gene promoters in 21 lung adenocarcinoma samples, as confirmed by bisulfite sequencing. We also developed a multiplex MS-FLAG assay for GATA5 and RASSF1 promoters. Conclusion: MS-FLAG provides a new, quantitative, high-throughput method for detecting gene promoter methylation and is a convenient alternative to agarose gel-based MSP for screening methylation. In addition to methylation, FLAG-based real-time signal generation may have broad applications in DNA diagnostics.
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Bruce, Sara, Katariina Hannula-Jouppi, Cecilia M. Lindgren, Marita Lipsanen-Nyman, and Juha Kere. "Restriction Site–Specific Methylation Studies of Imprinted Genes with Quantitative Real-Time PCR." Clinical Chemistry 54, no. 3 (March 1, 2008): 491–99. http://dx.doi.org/10.1373/clinchem.2007.098491.

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Abstract Background: Epigenetic studies, such as the measurement of DNA methylation, are important in the investigation of syndromes influenced by imprinted genes. Quick and accurate quantification of methylation at such genes can be of appreciable diagnostic aid. Methods: We first digested genomic DNA with methylation-sensitive restriction enzymes and used DNA without digestion as a control and nonmethylated λ DNA as an internal control for digestion efficiency. We then performed quantitative real-time PCR analyses with 6 unique PCR assays to investigate 4 imprinting control regions on chromosomes 7 and 11 in individuals with uniparental disomy of chromosome 7 (UPD7) and in control individuals. Results: Our validation of the method demonstrated both quantitative recovery and low methodologic imprecision. The imprinted loci on chromosome 7 behaved as expected in maternal UPD7 (100% methylation) and paternal UPD7 (<10% methylation). In controls, the mean (SD) for percent methylation at 2 previously well-studied restriction sites were 46% (6%) for both H19 and KCNQ1OT1, a result consistent with the previously observed methylation rate of approximately 50%. The methylation percentages of all investigated imprinted loci were normally distributed, implying that the mean and SD can be used as a reference for screening methylation loss or gain. Conclusion: The investigated loci are of particular importance for investigating the congenital Silver–Russell and Beckwith–Wiedemann syndromes; however, the method can also be applied to other imprinted regions. This method is easy to set up, has no PCR bias, requires small amounts of DNA, and can easily be applied to large patient populations for screening the loss or gain of methylation.
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Yoshioka, Masaki, Tomoo Matsutani, Ayaka Hara, Seiichiro Hirono, Takaki Hiwasa, Masaki Takiguchi, and Yasuo Iwadate. "Real-time methylation-specific PCR for the evaluation of methylation status of MGMT gene in glioblastoma." Oncotarget 9, no. 45 (June 12, 2018): 27728–35. http://dx.doi.org/10.18632/oncotarget.25543.

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Mehta, Akanksha, Anna Mielnik, DariusA Paduch, and PeterN Schlegel. "Novel methylation specific real-time PCR test for the diagnosis of Klinefelter syndrome." Asian Journal of Andrology 16, no. 5 (2014): 684. http://dx.doi.org/10.4103/1008-682x.125914.

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Vlassenbroeck, Ilse, Stéphane Califice, Annie-Claire Diserens, Eugenia Migliavacca, Josef Straub, Ivano Di Stefano, Fabrice Moreau, et al. "Validation of Real-Time Methylation-Specific PCR to Determine O6-Methylguanine-DNA Methyltransferase Gene Promoter Methylation in Glioma." Journal of Molecular Diagnostics 10, no. 4 (July 2008): 332–37. http://dx.doi.org/10.2353/jmoldx.2008.070169.

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Kondo, Masahiro, Hirofumi Aboshi, Masaaki Yoshikawa, Ayano Ogata, Ryosuke Murayama, Masami Takei, and Shin Aizawa. "A newly developed age estimation method based on CpG methylation of teeth-derived DNA using real-time methylation-specific PCR." Journal of Oral Science 63, no. 1 (2021): 54–58. http://dx.doi.org/10.2334/josnusd.20-0138.

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Kristensen, L. S., T. Mikeska, M. Krypuy, and A. Dobrovic. "Sensitive Melting Analysis after Real Time- Methylation Specific PCR (SMART-MSP): high-throughput and probe-free quantitative DNA methylation detection." Nucleic Acids Research 36, no. 7 (February 16, 2008): e42-e42. http://dx.doi.org/10.1093/nar/gkn113.

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Kristensen, Lasse Sommer, and Lise Lotte Hansen. "PCR-Based Methods for Detecting Single-Locus DNA Methylation Biomarkers in Cancer Diagnostics, Prognostics, and Response to Treatment." Clinical Chemistry 55, no. 8 (August 1, 2009): 1471–83. http://dx.doi.org/10.1373/clinchem.2008.121962.

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Abstract Background: DNA methylation is a highly characterized epigenetic modification of the human genome that is implicated in cancer. The altered DNA methylation patterns found in cancer cells include not only global hypomethylation but also discrete hypermethylation of specific genes. In particular, numerous tumor suppressor genes undergo epigenetic silencing because of hypermethylated promoter regions. Some of these genes are considered promising DNA methylation biomarkers for early cancer diagnostics, and some have been shown to be valuable for predicting prognosis or the response to therapy. Content: PCR-based methods that use sodium bisulfite–treated DNA as a template are generally accepted as the most analytically sensitive and specific techniques for analyzing DNA methylation at single loci. A number of new methods, such as methylation-specific fluorescent amplicon generation (MS-FLAG), methylation-sensitive high-resolution melting (MS-HRM), and sensitive melting analysis after real-time methylation-specific PCR (SMART-MSP), now complement the traditional PCR-based methods and promise to be valuable diagnostic tools. In particular, the HRM technique shows great potential as a diagnostic tool because of its closed-tube format and cost-effectiveness. Summary: Numerous traditional and new PCR-based methods have been developed for detecting DNA methylation at single loci. All have characteristic advantages and disadvantages, particularly with regard to use in clinical settings.
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Dissertations / Theses on the topic "Real-time methylation-specific PCR"

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Du, Toit Jean. "An analysis of the effect of transformation on global– and gene–specific DNA methylation in four cultured cell lines / Jean du Toit." Thesis, North-West University, 2010. http://hdl.handle.net/10394/4560.

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DNA methylation plays a role in several biological functions, such as gene expression regulation, and several endogenous and exogenous factors affect these DNA methylation patterns in the cell. One such alteration of a cell line's DNA methylation pattern is caused by the insertion of a vector into the cell line. Using the cytosine–extension assay and realtime methylation–specific PCR, alterations of DNA methylation levels on both global and gene–specific levels were investigated. In some cell lines the cellular transformation led to an increase in DNA methylation levels, and in others a decrease in DNA methylation amounts was observed. The same phenomenon was seen in the promoter regions of specific genes, showing that vector–insertion into a cell line caused DNA methylation alterations in many regions of the genome. These alterations in DNA methylation are investigated in this reduced representation study using enrichment of the methylated fraction of fragmented DNA and subsequent GS FLX Titanium sequencing of these methylated fragments. The results of sequence data analysis showed that methylated fragments are distributed over the whole genome, but could be related to only a few specific genes. These results have implications for cell culture work, biotechnological applications and uses in gene therapy.
Thesis (M.Sc. (Biochemistry))--North-West University, Potchefstroom Campus, 2011.
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Book chapters on the topic "Real-time methylation-specific PCR"

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Dugast-Darzacq, Claire, and Thierry Grange. "MethylQuant: A Real-Time PCR-Based Method to Quantify DNA Methylation at Single Specific Cytosines." In Methods in Molecular Biology, 281–303. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-59745-522-0_21.

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Conference papers on the topic "Real-time methylation-specific PCR"

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Riggs, Bridget, Amber Delmas, Kevin Brown, and Erin M. Siegel. "Abstract 4930: Comparing the methylation status in cervical squamous intraepithelial lesions using quantitative real-time methylation specific PCR (QMSP) and pyrosequencing." In Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1538-7445.am10-4930.

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Chung, Woonbok, Jolanta Bondaruk, Daniel D. Kim, Bogdan Czerniak, and Jean-Pierre J. Issa. "Abstract 4908: Detection of bladder tumor by quantitative methylation specific real time PCR from urine sediment DNA." In Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC. American Association for Cancer Research, 2010. http://dx.doi.org/10.1158/1538-7445.am10-4908.

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