Efficient recovery of environmental DNA for expression cloning by indirect extraction methodsGabor, E., de Vries, E. & Janssen, DB., 15-May-2003, In : FEMS Microbial Ecology. 44, 2, p. 153-163 11 p.
Research output: Contribution to journal › Article › Academic › peer-review
Using direct and cell extraction-based (indirect) isolation methods, DNA was obtained from environmental samples with largely differing characteristics (loam soil, sand soil, sediment, activated sludge, and compost) and evaluated with respect to the comprised bacterial diversity and its suitability for expression cloning in Escherichia coli. Indirect DNA extraction methods yielded 10 to 100-fold lower amounts of DNA than direct procedures, but the bacterial diversity of DNA recovered by indirect means was distinctly higher as shown by denaturing gradient gel electrophoresis. Furthermore, much lower amounts of eukaryotic DNA were co-extracted if cell extraction-based methods were used (
|Number of pages||11|
|Journal||FEMS Microbial Ecology|
|Publication status||Published - 15-May-2003|
- expression cloning, denaturing gradient gel electrophoresis, environmental gene bank, cell extraction, direct DNA isolation, GRADIENT GEL-ELECTROPHORESIS, 16S RIBOSOMAL-RNA, POLYMERASE-CHAIN-REACTION, MICROBIAL DIVERSITY, ESCHERICHIA-COLI, BACTERIAL COMMUNITY, SEDIMENT SAMPLES, GENOMIC DNA, SOIL, LIBRARIES