Jackson D P, Lewis F A, Taylor G R, Boylston A W, Quirke P
Department of Pathology, University of Leeds.
J Clin Pathol. 1990 Jun;43(6):499-504. doi: 10.1136/jcp.43.6.499.
Several DNA extraction techniques were quantitatively and qualitatively compared using both fresh and paraffin wax embedded tissue and their suitability investigated for providing DNA and RNA for the polymerase chain reaction (PCR). A one hour incubation with proteinase K was the most efficient DNA extraction procedure for fresh tissue. For paraffin wax embedded tissue a five day incubation with proteinase K was required to produce good yields of DNA. Incubation with sodium dodecyl sulphate produced very poor yields, while boiling produced 20% as much DNA as long enzyme digestion. DNA extracted by these methods was suitable for the PCR amplification of a single copy gene. Proteinase K digestion also produced considerable amounts of RNA which has previously been shown to be suitable for PCR analysis. A delay before fixation had no effect on the amount of DNA obtained while fixation in Carnoy's reagent results in a much better preservation of DNA than formalin fixation, allowing greater yields to be extracted.
使用新鲜组织和石蜡包埋组织对几种DNA提取技术进行了定量和定性比较，并研究了它们为聚合酶链反应（PCR）提供DNA和RNA的适用性。用蛋白酶K孵育1小时是从新鲜组织中提取DNA最有效的方法。对于石蜡包埋组织，需要用蛋白酶K孵育5天才能获得高产率的DNA。用十二烷基硫酸钠孵育产生的产量非常低，而煮沸产生的DNA量是长时间酶消化的20%。通过这些方法提取的DNA适用于单拷贝基因的PCR扩增。蛋白酶K消化还产生了大量RNA，此前已证明这些RNA适用于PCR分析。固定前的延迟对获得的DNA量没有影响，而用卡诺氏试剂固定比福尔马林固定能更好地保存DNA，从而可以提取更高的产量。
