Jadaon Mehrez M, Dashti Ali A, Lewis Hend L, Habeeb Fatema M
Department of Medical Laboratory Sciences, Faculty of Allied Health Sciences, Kuwait University, Sulaibekhat 90805, Kuwait.
Med Princ Pract. 2009;18(4):280-3. doi: 10.1159/000215724. Epub 2009 Jun 2.
The aim of the present study was to develop a simple, quick and cheap method to process whole-blood samples for the molecular techniques polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) without the use of expensive reagents or sophisticated machines.
Venous whole-blood samples were collected from 40 individuals. The samples were frozen at -80 degrees C, and then rapidly thawed at 37 degrees C. Each sample was incubated with distilled water, then boiled in a microwave and centrifuged. The supernatant was taken directly for PCR and RFLP. For comparison, PCR and RFLP were performed on DNA purified from the same samples using the phenol-chloroform method and two commercial DNA extraction kits.
PCR/RFLP results using the presented method were qualitatively similar to those obtained by DNA extracted using the other three methods.
The presented method proved to be a simpler and cheaper way of processing whole-blood samples for PCR and RFLP analyses.
本研究的目的是开发一种简单、快速且廉价的方法，用于处理全血样本以进行聚合酶链反应（PCR）和限制性片段长度多态性（RFLP）等分子技术，而无需使用昂贵的试剂或精密仪器。
从40名个体采集静脉全血样本。样本在-80℃冷冻，然后在37℃快速解冻。每个样本用蒸馏水孵育，然后在微波炉中煮沸并离心。直接取上清液进行PCR和RFLP。为作比较，使用酚-氯仿法和两种商用DNA提取试剂盒，对从相同样本中纯化的DNA进行PCR和RFLP。
使用本方法获得的PCR/RFLP结果在质量上与使用其他三种方法提取DNA所获得的结果相似。
对于PCR和RFLP分析而言，本方法被证明是一种更简单、更廉价的处理全血样本的方法。
