Empirical evaluation of preservation methods for faecal DNA

被引:209
作者
Frantzen, MAJ
Silk, JB
Ferguson, JWH
Wayne, RK
Kohn, MH [1 ]
机构
[1] Univ Calif Los Angeles, Dept Biol, Los Angeles, CA 90095 USA
[2] Univ Pretoria, Dept Zool & Entomol, ZA-0001 Pretoria, South Africa
[3] Univ Calif Los Angeles, Dept Anthropol, Los Angeles, CA 90095 USA
关键词
apolipoprotein E; cytochrome c oxidase II; DNA preservation; molecular scatology; noninvasive sampling; Papio cynocephalus ursinus;
D O I
10.1046/j.1365-294x.1998.00449.x
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
We evaluate the relative effectiveness of four methods for preserving faecal samples for DNA analysis. PCR assays of fresh faecal samples collected from free-ranging baboons showed that amplification success was dependent on preservation method, PCR-product size, and whether nuclear or mitochondrial DNA was assayed. Storage in a DMSO/EDTA/Tris/salt solution (DETs) was most effective for preserving nuclear DNA, but storage in 70% ethanol, freezing at -20 degrees C and drying performed approximately equally well for mitochondrial DNA and short (<200 bp) nuclear DNA fragments. Because faecal DNA is diluted and degraded, repeated extractions from faeces may be necessary and short nuclear markers should be employed for genotyping. A review of molecular scatology studies further suggests that three to six faeces per individual should be collected.
引用
收藏
页码:1423 / 1428
页数:6
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