変性剤濃度勾配ゲル電気泳動法による微生物群集プロファイリングの有効性
書誌事項
- タイトル別名
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- Quantitative validity of microbial community profiling by denaturing gradient gel electrophoresis.
抄録
Using known 16S ribosomal DNA fragments amplified by PCR from different bacteria, we evaluated the validity of denaturing gradient gel electrophoresis (DGGE) profiling as a quantitative method to estimate the microbial community in freshwater ecosystems. The number of copies as template DNAs was 104-105 in 100μL of 30-cycle PCR reaction to obtain a detectable DGGE band stained with ethidium bromide under transillumina-tion. Moreover, we compared the band abundances on the DGGE profiling with the band mixture of 16S rDNA fragments blended in PCR solution to evaluate whether DGGE band intensities reflect true gene abundance. As the result, the optimum copy number of mixed template DNA fragments was 105 order in 100μL of PCR solution to estimate the accurate the ratio of mixed DNA by PCR-DGGE. As the abundance of one template DNA was more than 86% of total template DNAs in the PCR reaction solution, the difference in the ratio between the abundance of template DNA and that of DGGE band intensity was decreased. These results indicate that it is important to adjust the concentration of template DNA in the PCR reaction solution in order to perform the PCR-DGGE analysis for evaluation of a predominant microorganism in the microbial community.
収録刊行物
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- 陸水学雑誌
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陸水学雑誌 63 (1), 59-66, 2002
日本陸水学会
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詳細情報 詳細情報について
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- CRID
- 1390282679036003584
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- NII論文ID
- 130003645136
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- ISSN
- 18824897
- 00215104
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- 本文言語コード
- ja
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- データソース種別
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- JaLC
- Crossref
- CiNii Articles
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- 抄録ライセンスフラグ
- 使用不可