Hundredfold productivity of genome analysis by introduction of microtemperature-gradient gel electrophoresis.
Genome profiling, which employs temperature-gradient gel electrophoresis (TGGE) for DNA analysis, has recently been developed in identifying species by genotype. However, the performance of this technology like the general applications of TGGE was, though highly informative, limited in its ability due to methodological reasons. This study demonstrates that minimization of the gel for TGGE, to around one-tenth of its conventional size (approximately 2 cm), can be successfully introduced, resulting in a hundredfold higher performance (total evaluation of time, cost, and degree of parallel operations) than that of the conventional. Reproducibility was evaluated from the measures of the pattern similarity scores (PaSS) between band patterns (genome profiles) obtained with the conventional TGGE, and that with micro-TGGE (microTGGE) developed here, after extracting a set of featuring points from genome profiles. Size minimization, which leads to the reduction of the amount of samples required (cost-saving), is another great advantage, enhancing the employment of multicolor fluorescence technology. Since the further development of microbe-related fields such as epidemiology and microbial ecology inevitably require knowledge based on the identification of a great number of species and strains, microbe-related fields will receive the most optimal benefits from the technological improvements attained here.
Biyani
M
M
Department of Functional Materials Science, Saitama University, Urawa, Saitama, Japan.
Nishigaki
K
K
D001407:Bacillus / Q000235:genetics; D001412:Bacillus subtilis / Q000235:genetics; D004269:DNA, Bacterial / Q000032:analysis*; D004271:DNA, Fungal / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics; D016680:Genome, Bacterial; D016681:Genome, Fungal; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013194:Staining and Labeling / Q000379:methods; D013696:Temperature
D004269:DNA, Bacterial; D004271:DNA, Fungal