Genomic DNA from Mice: A Comparison of Recovery Methods and Tissue Sources

John R. Hofstetter , Aiwu Zhang , Aimee R. Mayeda , Tim Guscar , John I. Nurnberger Jr. , Debomoy K. Lahiri
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引用次数: 55

Abstract

Our aim is to identify an extraction method and the source of mouse tissue(s) that could allow a high-resolution genomic scan from a living mouse. We compared and optimized two methods for yield, purity of DNA, and their use in the polymerase chain reaction (PCR) of DNA extracted from different mouse tissues. In addition to whole blood, tissue samples from the brain, liver, testis, and tail were included in this study. The Rapid Method (RM) is preferable for the whole blood samples and testis and brain tissue samples because it is quicker, less toxic, and more cost-effective than the proteinase K method (PM). For liver the PM produced higher yields of DNA with less degradation than the RM. For tail tip, the PM produced a higher yield of DNA, but the RM resulted in a higher yield of PCR product. From a living mouse, a tail snip generated a sufficient amount of DNA for several hundred PCRs but not a complete genomic scan. We suggest that the RM can be used to extract genomic DNA for a complete genomic scan which requires either testicular tissues or repeated blood samples from the suborbital sinus over several months without sacrificing the animal.

小鼠基因组DNA:恢复方法和组织来源的比较
我们的目标是确定一种提取方法和小鼠组织的来源,从而可以对活体小鼠进行高分辨率的基因组扫描。我们比较和优化了两种方法的产量、DNA的纯度,以及它们在不同小鼠组织中提取DNA的聚合酶链反应(PCR)中的应用。除全血外,本研究还包括来自大脑、肝脏、睾丸和尾巴的组织样本。快速法(Rapid Method, RM)比蛋白酶K法(PM)更快、毒性更低、成本更低,更适合全血样本、睾丸和脑组织样本。对于肝脏,PM比RM产生更高的DNA产量和更少的降解。对于尾尖,PM产生更高的DNA产量,而RM产生更高的PCR产物产量。从一只活着的老鼠身上,一段尾巴剪断产生了足够的DNA,可以进行几百次pcr,但不能进行完整的基因组扫描。我们建议RM可用于提取基因组DNA进行完整的基因组扫描,这需要在几个月内从睾丸组织或从眶下窦重复采集血液样本,而不会牺牲动物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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