技法シリーズ
微生物生態学への変性剤濃度勾配ゲル電気泳動法の応用
2000 年 15 巻 1 号 p. 59-73
詳細
2000 年 15 巻 1 号 p. 59-73
The use of Denaturing Gradient Gel Electrophoresis(DGGE)has been widely recognized in microbial ecology and its importance has increased recently.DNA extracted from the environment is amplified by PCR using a set of primers in which one of the two contains the 40 base GC-rich sequence at the 5′ -end prior to DGGE. The PCR amplified DNA fragment migrates in a DGGE gel depending on its melting behavior. Moreover, the DNA resolved by DGGE can be promoted for further analyses, such as hybridization and sequencing without any cloning step. For these reasons, PCR-DGGE is one of the powerful tools available for the detection and phylogenetic analysis of environmental microbes. Although this method is easy and reproducible, some knowledge and technique are required to obtain reliable data. This technical series introduces protocols for the PCR-DGGE method with the theory presented to beginners as comprehensibly as possible. In addition, we discussed the application and limitation of the PCR-DGGE approach to microbial ecology.
