Noncryogenic preservation of mammalian tissues for DNA extraction: An assessment of storage methods
Noncryogenic preservation of mammalian tissues for DNA extraction: An assessment of storage methods
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DOI：
10.1023/a:1014541222816
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发表时间：
2002-02-01
影响因子：
2.4
通讯作者：
Kilpatrick, CW
中科院分区：
文献类型：
--
作者：
Kilpatrick, CW
用于DNA提取和扩增的组织的可靠的现场储存方法对于采用分子技术的许多研究至关重要。保护DNA降解进行了比较三种常用的方法，非低温储存的组织超过2年的时间尺度。所有三种方法在储存期间防止DNA降解至少6个月。DMSO（二甲基亚砜）盐溶液为储存长达2年的组织提供了最佳的DNA降解保护。从组织储存2年的裂解缓冲液中回收高分子量DNA，然而，也存在中等量的降解DNA。高分子量的DNA回收从组织中存储在乙醇中2年，然而，产量相对较小相比，其他两个noncryogenic存储技术。乙醇保存的组织中的大部分DNA降解似乎发生在提取过程中，并且可以通过在开始提取之前将组织浸泡在裂解缓冲液中几个小时来减少。从DMSO-盐溶液保存的组织中提取的DNA的PCR产物的产率是最大的，而从储存在裂解缓冲液或乙醇中的组织中提取的DNA的产率较低。
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 tit 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.