Tsai Y L, Olson B H
Environmental Design and Analysis, Program in Social Ecology, University of California, Irvine 92717.
Appl Environ Microbiol. 1991 Apr;57(4):1070-4. doi: 10.1128/aem.57.4.1070-1074.1991.
A rapid method for the direct extraction of DNA from soil and sediments was developed. The indigenous microorganisms in the soil and sediments were lysed by using lysozyme and a freeze-thaw procedure. The lysate was extracted with sodium dodecyl sulfate and phenol-chloroform. In addition to a high recovery efficiency (greater than 90%), the yields of DNA were high (38 and 12 micrograms/g [wet weight] from sediments and soil, respectively). This method generated minimal shearing of the extracted DNA. The crude DNA could be further purified with an Elutip-d column if necessary. An additional advantage of this method is that only 1 g of sample is required, which allows for the analysis of small samples and the processing of many samples in a relatively short (7 h) period.
开发了一种从土壤和沉积物中直接提取DNA的快速方法。通过使用溶菌酶和冻融程序裂解土壤和沉积物中的土著微生物。裂解物用十二烷基硫酸钠和苯酚-氯仿提取。除了具有较高的回收效率(大于90%)外,DNA产量也很高(沉积物和土壤分别为38和12微克/克[湿重])。该方法对提取的DNA造成的剪切最小。如有必要,粗DNA可用Elutip-d柱进一步纯化。该方法的另一个优点是仅需1克样品,这使得可以分析小样品,并在相对较短的时间(7小时)内处理多个样品。