Tuesday, September 10, 2019
Evaluate the methods used to obtain qualitative and quantitive Essay
Evaluate the methods used to obtain qualitative and quantitive information on DNA methylation - Essay Example Varied methods for analyzing DNA methylation quantitatively and qualitatively exist, and they entail studying the polymerase chain reactions. Notably, the identified and widely procedures for analyzing DNA methylation both qualitatively and quantitatively include methylation-specific PCR(MSP), combined bisulfite restriction analysis (COBRA), bisulfite sequencing, methylation-sensitive single nucleotide primer extension (Ms-SNuPE), as well as, quantitative real-time MSP (Szyf, 2010 p 29). The use of each technique depends on the efficacy and intended information gain of the analysis process. Methylation-specific PCR technique involves the analysis of DNA methylation patterns in CpG islands. Thus, it is a qualitative technique of obtaining information on DNA methylation. The method involves the modification of the DNA using PCR with two primer pairs that are detectable in methylated and unmethylated DNA respectively (Ip, Canham, Choo, Inaba, Jacobs, Kalitsis, Mattiske, Saffery, Wong, Wong & Mann, 2012, p 9). The study of the methylation patterns focuses on the regulation of gene expressions as they occur in mammalian cells. This technique is useful from its advantages. Notably, it is highly sensitive and highly specific for particulate CpG sites, which makes it remarkably accurate. Additionally, the Methylation-specific PCR (MSP) facilitates for analysis of clinical samples, which have low levels of methylated sequences. The use of the restriction enzymes eliminates the problem of incomplete enzyme digestion; hence, giving accurate findings. The limitation of the techniq ue includes that incomplete bisulfite modification of the DNA gives false positivity results (Szyf, 2010 p 56). Further, poor design of the primers could give inconclusive results. Bisulfite sequencing technique entails the use of bisulphite treatment of the DNA to establish its pattern of methylation. It is a qualitative technique and was the first
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