Open AccessObservation of bias associated with re‐amplification of DNA isolated from denaturing gradient gels
Author(s) -
Nikolausz Marcell,
Sipos Rita,
Révész Sára,
Székely Anna,
Márialigeti Károly
Publication year - 2005
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/j.femsle.2005.02.013
Subject(s) - temperature gradient gel electrophoresis , amplicon , gel electrophoresis , dna , biology , polymerase chain reaction , genetics , microbiology and biotechnology , gene , 16s ribosomal rna
DNA from environmental PCR products separated by denaturing gradient gel electrophoresis (DGGE) was isolated from the background smear rather than from discrete bands of the DGGE gel. The “interband” region was considered as a potential source of less dominant members of natural microbial communities. Surprisingly, instead of detecting new bands from the re‐amplified PCR products, patterns very similar to the original ones were obtained regardless of the position of the “interband” region. The results suggest that the separation of amplicons by DGGE may not be perfect and band re‐amplification based sequence analyses need careful interpretation.