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使用聚乙烯聚吡咯烷酮离心柱从土壤中快速直接提取用于聚合酶链反应(PCR)分析的DNA。

Rapid, direct extraction of DNA from soils for PCR analysis using polyvinylpolypyrrolidone spin columns.

作者信息

Berthelet M, Whyte L G, Greer C W

机构信息

National Research Council of Canada, Biotechnology Research Institute, Montréal, Québec, Canada.

出版信息

FEMS Microbiol Lett. 1996 Apr 15;138(1):17-22. doi: 10.1111/j.1574-6968.1996.tb08128.x.

Abstract

Polyvinylpolypyrrolidone spin columns were used to rapidly purify crude soil DNA extracts from humic materials for polymerase chain reaction (PCR) analysis. The PCR detection limit for the tfdC gene, encoding chlorocatechol dioxygenase from the 2,4-dichlorophenoxyacetic acid degradation pathway, was 10(1)-10(2) cells/g soil in inoculated soils. The procedure could be applied to the amplification of biodegradative genes from indigenous microbial populations from a wide variety of soil types, and the entire analysis could be performed within 8 h.

摘要

聚乙烯聚吡咯烷酮旋转柱用于从腐殖质中快速纯化粗制土壤DNA提取物,以进行聚合酶链反应(PCR)分析。编码来自2,4-二氯苯氧乙酸降解途径的氯儿茶酚双加氧酶的tfdC基因的PCR检测限在接种土壤中为10(1)-10(2)个细胞/克土壤。该方法可应用于从多种土壤类型的本地微生物群体中扩增生物降解基因,整个分析可在8小时内完成。

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