Denaturing gradient gel electrophoresis gel expansion (DGGEGE)--an attempt to resolve the limitations of co-migration in the DGGE of complex polymicrobial communities.
Recent molecular approaches for the study of microbial communities such as PCR-cloning have enabled the detection and identification of as-yet-unculturable taxa. Cloning and sequencing of multiple samples is extremely laborious and expensive to perform thoroughly due to the large diversity involved. For this purpose, techniques such as denaturing gradient gel electrophoresis (DGGE) may be better suited. There is increasing evidence suggesting that DGGE of complex polymicrobial communities may be limited by co-migration of different sequences. In this study, we attempt to address this limitation by excising individual bands and running them through a shorter denaturant gradient, a process we have termed "denaturing gradient gel electrophoresis gel expansion" (DGGEGE).
Gafan
Gavin P
GP
Division of Microbial Diseases, UCL Eastman Dental Institute, 256 Gray's Inn Road, London WC1X 8LD, UK.
Spratt
David A
DA
D001419:Bacteria / Q000145:classification* / Q000235:genetics; D044822:Biodiversity; D002648:Child; D002675:Child, Preschool; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D003773:Dental Plaque / Q000382:microbiology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D014644:Genetic Variation; D006801:Humans; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D004269:DNA, Bacterial