DETECTION OF LOW NUMBERS OF BACTERIAL-CELLS IN SOILS AND SEDIMENTS BY POLYMERASE CHAIN-REACTION
DETECTION OF LOW NUMBERS OF BACTERIAL-CELLS IN SOILS AND SEDIMENTS BY POLYMERASE CHAIN-REACTION
复制标题
DOI：
10.1128/aem.58.2.754-757.1992
复制
发表时间：
1992-02-01
影响因子：
4.4
通讯作者：
OLSON, BH
中科院分区：
文献类型：
--
作者：
TSAI, YL;OLSON, BH
采用聚合酶链反应对土壤和沉积物提取物中两个低拷贝数的16S核糖体基因片段进行扩增。用快速冻融法从1g有籽或未籽样品中提取用于聚合酶链反应的总DNA。从含有少于3个大肠杆菌细胞的种子土壤和含有少于10个细胞的种子沉积物中分离的DNA片段可以检测到扩增的DNA片段。该研究表明，将聚合酶链反应与直接DNA提取相结合，对沉积物和土壤的灵敏度分别提高了1个和2个数量级。这项技术可能成为遗传生态学研究的有力工具。
Polymerase chain reaction was used to amplify the low copy number of two 16S ribosomal gene fragments from soil and sediment extracts. Total DNA for polymerase chain reaction was extracted from 1 g of seeded or unseeded samples by a rapid freeze-and-thaw method. Amplified DNA fragments can be detected in DNA fractions isolated from seeded soil containing less than 3 Escherichia coli cells and from seeded sediments containing less than 10 cells. This research demonstrated that coupling polymerase chain reaction to direct DNA extraction improves sensitivity by 1 and 2 orders of magnitude for sediments and soils, respectively. This technique could become a powerful tool for genetic ecology studies.