Kilpatrick C William
Department of Biology, University of Vermont, Burlington 05405-0086, USA.
Biochem Genet. 2002 Feb;40(1-2):53-62. doi: 10.1023/a:1014541222816.
Reliable field methods for the storage of tissues to be used for DNA extraction and amplification are critical to many studies employing molecular techniques. Protection from DNA degradation was compared among three commonly used methods of noncryogenic storage of tissues over a time scale of 2 years. All three methods prevented DNA degradation during storage for at least 6 months. DMSO (dimethyl sulfoxide)-salt solution provided the best protection from DNA degradation of tissues stored for up to 2 years. High molecular weight DNA was recovered from lysis buffer in which tissue was stored for 2 years, however, moderate amounts of degraded DNA was also present. High molecular weight DNA was recovered from tissues stored in ethanol for 2 years, however the yield was relatively small compared to the other two noncryogenic storage techniques. Much of the DNA degradation in ethanol preserved tissues appeared to occur during the extraction procedure and can be reduced by soaking the tissue in lysis buffer for a few hours prior to beginning the extraction. The yield of PCR products was greatest from DNA extracted from DMSO-salt solution preserved tissues, whereas DNA from tissues stored in either lysis buffer or ethanol produced lower yields.
对于许多采用分子技术的研究而言，用于DNA提取和扩增的组织储存的可靠现场方法至关重要。在两年的时间范围内，比较了三种常用的非低温组织储存方法对DNA降解的保护作用。所有三种方法在储存至少6个月期间都能防止DNA降解。二甲基亚砜（DMSO）-盐溶液对储存长达2年的组织提供了最佳的DNA降解保护。从储存2年的组织的裂解缓冲液中回收了高分子量DNA，然而，也存在中等量的降解DNA。从在乙醇中储存2年的组织中回收了高分子量DNA，但是与其他两种非低温储存技术相比，产量相对较小。乙醇保存的组织中的许多DNA降解似乎发生在提取过程中，并且可以通过在开始提取前将组织在裂解缓冲液中浸泡几个小时来减少。从DMSO-盐溶液保存的组织中提取的DNA的PCR产物产量最高，而从储存在裂解缓冲液或乙醇中的组织中提取的DNA产量较低。
