Nikolausz Marcell, Sipos Rita, Révész Sára, Székely Anna, Márialigeti Károly
Department of Microbiology, Eötvös Loránd University of Science, Pázmány Péter sétány 1/c, 1117 Budapest, Hungary.
FEMS Microbiol Lett. 2005 Mar 15;244(2):385-90. doi: 10.1016/j.femsle.2005.02.013.
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.
通过变性梯度凝胶电泳（DGGE）分离的环境PCR产物中的DNA是从背景条带中分离出来的，而非从DGGE凝胶的离散条带中分离得到。“条带间”区域被认为是天然微生物群落中不太占优势成员的潜在来源。令人惊讶的是，重新扩增的PCR产物并未检测到新的条带，无论“条带间”区域的位置如何，得到的条带模式都与原始模式非常相似。结果表明，DGGE对扩增子的分离可能并不完美，基于条带重新扩增的序列分析需要谨慎解读。
