Noncryogenic preservation of mammalian tissues for DNA extraction: An assessment of storage methods

被引:75
作者
Kilpatrick, CW [1 ]
机构
[1] Univ Vermont, Dept Biol, Burlington, VT 05405 USA
关键词
tissue storage; noncryogenic; DNA extraction; degradation;
D O I
10.1023/A:1014541222816
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:53 / 62
页数:10
相关论文
共 17 条
[1]   COMPARATIVE STUDIES OF AVIAN DNA BY RESTRICTION FRAGMENT LENGTH POLYMORPHISM ANALYSIS - CONVENIENT PROCEDURES ON BLOOD-SAMPLES FROM LIVE BIRDS [J].
ARCTANDER, P .
JOURNAL FUR ORNITHOLOGIE, 1988, 129 (02) :205-216
[2]   GENERAL METHOD FOR ISOLATION OF HIGH MOLECULAR-WEIGHT DNA FROM EUKARYOTES [J].
BLIN, N ;
STAFFORD, DW .
NUCLEIC ACIDS RESEARCH, 1976, 3 (09) :2303-2308
[3]  
COCKBURN AF, 1988, J AM MOSQUITO CONTR, V4, P261
[4]   PHARMACOLOGY OF DIMETHYL SULFOXIDE [J].
DAVID, NA .
ANNUAL REVIEW OF PHARMACOLOGY, 1972, 12 :353-+
[5]  
DESSAUER HC, 1984, COLLECTIONS FROZEN T, P74
[6]  
Dessauer Herbert C., 1996, P29
[7]  
Freitas-Sibajev Maria Goreti, 1995, Mosquito Systematics, V27, P78
[8]  
HOUDE P, 1988, AUK, V105, P773
[9]  
Longmire Jonathan L., 1997, Occasional Papers the Museum Texas Tech University, V163, P1
[10]  
MASON MM, 1971, DIMETHYL SULFOXIDE, P113