Gustafson S, Proper J A, Bowie E J, Sommer S S
Department of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905.
Anal Biochem. 1987 Sep;165(2):294-9. doi: 10.1016/0003-2697(87)90272-7.
An examination of variables affecting the yield of DNA from blood was undertaken in order to improve sample processing and to evaluate alternative methods of mailing blood samples for DNA analysis. A rapid, high-yield method was developed for the isolation of high-molecular-weight DNA from fresh and frozen blood. In addition, the following observations were made: (1) Of the anticoagulants examined, acid citrate dextrose (ACD) solution B was found to be superior to EDTA and heparin for preserving yields of DNA during incubation at room temperature. If DNA is isolated from frozen blood, high yields of undegraded DNA are achieved after incubation at 23 degrees C for 5 days with ACD solution B. (2) High yields of undegraded DNA are obtained from blood stored with ACD solution B for at least 1 day at 42 degrees C, 5 days at 0 degrees C, or 1 month at -20 degrees C. (3) Three cycles of freezing and thawing may have little if any affect on the yield of DNA. The results indicate that blood for DNA extraction may be mailed in an ambient temperature container and, in many cases, sent by first-class mail rather than by overnight delivery services.
为了改进样本处理方法并评估邮寄血样进行DNA分析的替代方法,我们对影响从血液中提取DNA产量的变量进行了研究。我们开发了一种快速、高产的方法,用于从新鲜血液和冷冻血液中分离高分子量DNA。此外,还得出了以下观察结果:(1)在所检测的抗凝剂中,发现酸性枸橼酸盐葡萄糖(ACD)溶液B在室温孵育期间保存DNA产量方面优于乙二胺四乙酸(EDTA)和肝素。如果从冷冻血液中分离DNA,在23摄氏度下用ACD溶液B孵育5天后可获得高产率的未降解DNA。(2)从在42摄氏度下用ACD溶液B储存至少1天、在0摄氏度下储存5天或在 -20摄氏度下储存1个月的血液中可获得高产率的未降解DNA。(3)三个冻融循环对DNA产量的影响可能很小或没有影响。结果表明,用于DNA提取的血液可以装在常温容器中邮寄,并且在许多情况下,可以通过一类邮件寄送,而不是通过隔夜快递服务。