Southern Blotting Application – Article Example
Southern Blotting Application A study by Taylor et al. (343) used Southern blotting technique to compare its sensitivitywith PCR amplification in detecting the germ line minisatellite mutation (GMM) that occur at the CEB1 locus. Taylor et al. utilized Southern hybridization in performing analysis of unamplified DNA. This entailed the use of minisatellite probes. The use of Southern blotting was to help the researchers make a complete comparison between these two techniques. Taylor et al. point that detection of GMM by Southern blotting is necessary for performing a number of epidemiological studies regarding the effects of genotoxins and radiations. The amount of DNA required per sample for Southern blot was comparatively larger than the one needed for the PCR amplification (Taylor et al., 346). Thus, Taylor et al. pointed that PCR was more sensitive than Southern blot.
PCR and Southern blot GMM assays vary concerning the size of alleles detected. Taylor and colleagues demonstrated that Southern blot technique could detect CEB1 alleles that were greater than 8.5kb, which PCR amplification could not. Thus, it is essential to point that Southern blot is sensitive to a large amount of DNA, whereas PCR amplification is sensitive to a small amount of DNA. One advantage of Southern blot technique, which the researchers found out is that using different probes it can screen by stripping and re-hybridizing blots of GMM at several loci, which PCR could not. Based on their empirical results, Taylor et al. concluded that PCR and Southern blot GMM analysis showed similar specificities; however, PCR amplification demonstrated more sensitivity with smaller DNA than Southern blot technique.
Taylor, Malcolm et al. “Comparison of Germ Line Minisatellite Mutation Detection at the CEB1 Locus by Southern Blotting and PCR Amplification.” Mutagenesis 25.4 (2010): 343–349. PubMed Central. Web. 19 Mar. 2015.