| RRC ID |
19077
|
| 著者 |
Kouduka M, Suko T, Morono Y, Inagaki F, Ito K, Suzuki Y.
|
| タイトル |
A new DNA extraction method by controlled alkaline treatments from consolidated subsurface sediments.
|
| ジャーナル |
FEMS Microbiol Lett
|
| Abstract |
Microbial communities that thrive in subterranean consolidated sediments are largely unknown owing to the difficulty of extracting DNA. As this difficulty is often attributed to DNA binding onto the silica-bearing sediment matrix, we developed a DNA extraction method for consolidated sediment from the deep subsurface in which silica minerals were dissolved by being heated under alkaline conditions. NaOH concentrations (0.07 and 0.33 N), incubation temperatures (65 and 94 °C) and incubation times (30-90 min) before neutralization were evaluated based on the copy number of extracted prokaryotic DNA. Prokaryotic DNA was detected by quantitative PCR analysis after heating the sediment sample at 94 °C in 0.33 N NaOH solution for 50-80 min. Results of 16S rRNA gene sequence analysis of the extracted DNA were all consistent with regard to the dominant occurrence of the metallophilic bacterium, Cupriavidus metallidurans, and Pseudomonas spp. Mineralogical analysis revealed that the dissolution of a silica mineral (opal-CT) during alkaline treatment was maximized at 94 °C in 0.33 N NaOH solution for 50 min, which may have resulted in the release of DNA into solution. Because the optimized protocol for DNA extraction is applicable to subterranean consolidated sediments from a different locality, the method developed here has the potential to expand our understanding of the microbial community structure of the deep biosphere.
|
| 巻・号 |
326(1)
|
| ページ |
47-54
|
| 公開日 |
2012-1-1
|
| DOI |
10.1111/j.1574-6968.2011.02437.x
|
| PMID |
22092362
|
| MeSH |
Cupriavidus / genetics
Cupriavidus / isolation & purification*
DNA, Bacterial / analysis
DNA, Bacterial / genetics
DNA, Bacterial / isolation & purification*
DNA, Ribosomal / analysis
DNA, Ribosomal / genetics
Genes, rRNA
Geologic Sediments / microbiology*
Molecular Sequence Data
Pseudomonas / genetics
RNA, Ribosomal, 16S / genetics
Silicon Dioxide
|
| IF |
1.987
|
| 引用数 |
12
|
|
WOS 分野
|
MICROBIOLOGY
|
| リソース情報 |
| 一般微生物 |
JCM 5965 |
