Factors affecting mitochondrial DNA quality from museum preserved Drosophila simulans
Factors affecting mitochondrial DNA quality from museum preserved Drosophila simulans
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DOI：
10.1023/a:1018963131302
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发表时间：
2001-03-01
影响因子：
1.9
通讯作者：
Ballard, JWO
中科院分区：
文献类型：
--
作者：
Dean, MD;Ballard, JWO
在这项研究中，我们调查了传统的杀死方法和储存制度如何影响拟果蝇线粒体DNA质量。在这里，我们用三个标准来定义质量：(1)提取DNA的大小，(2)提取率，(3)从四个目标区域扩增的能力。杀灭方法对提取率有显著影响，但对PCR成功率无显著影响。暴露于氰化物的标本中提取的DNA产量最高，而在70%乙醇中杀死的标本中提取的DNA产量最低。保存两年的标本含有严重剪切的DNA，与新检测的标本相比，这导致PCR成功率显著降低。与959 bp和291 bp片段相比，1822 bp和1332 bp片段的PCR成功率下降最为明显。萘不影响DNA质量的任何方面；无论在何种环境下，储存时间都会影响PCR的成功。这项研究有助于进一步完善我们对DNA降解的理解。
In this study we investigate how traditional killing methods and storage regimes affected mitochondrial DNA quality in Drosophila simulans. Here we define quality with three criteria: (1) size of extracted DNA, (2) extraction yield, and (3) ability to amplify from four target regions. Killing methods had a significant effect on extraction yield, but not on PCR success. Highest DNA yields were extracted from specimens exposed to cyanide, while the lowest were from specimens killed in 70% ethanol. Specimens stored for two years contained badly sheared DNA, which translated into a significant decrease in PCR success compared to freshly assayed specimens. The most dramatic decrease in PCR success occurred in the 1822 bp and 1332 bp amplicons, compared to the 959 bp and 291 bp fragments. Naphthalene did not affect any aspect of DNA quality; time of storage affected PCR success regardless of naphthalene environment. This study serves to further refine our understanding of DNA degradation.