一种用于提高从单个蚊子中提取DNA产量的DNA提取方案。
A DNA extraction protocol for improved DNA yield from individual mosquitoes.
作者信息
Nieman Catelyn C, Yamasaki Youki, Collier Travis C, Lee Yoosook
机构信息
Vector Genetics Laboratory, Department of Pathology, Microbiology, and Immunology, University of California, Davis, CA, USA.
出版信息
F1000Res. 2015 Nov 20;4:1314. doi: 10.12688/f1000research.7413.1. eCollection 2015.
Typical DNA extraction protocols from commercially available kits provide an adequate amount of DNA from a single individual mosquito sufficient for PCR-based assays. However, next-generation sequencing applications and high-throughput SNP genotyping assays exposed the limitation of DNA quantity one usually gets from a single individual mosquito. Whole genome amplification could alleviate the issue but it also creates bias in genome representation. While trying to find alternative DNA extraction protocols for improved DNA yield, we found that a combination of the tissue lysis protocol from Life Technologies and the DNA extraction protocol from Qiagen yielded a higher DNA amount than the protocol using the Qiagen or Life Technologies kit only. We have not rigorously tested all the possible combinations of extraction protocols; we also only tested this on mosquito samples. Therefore, our finding should be noted as a suggestion for improving people's own DNA extraction protocols and not as an advertisement of a commercially available product.
市售试剂盒的典型DNA提取方案能从单个蚊子个体中提供足够量的DNA，足以用于基于PCR的检测。然而，新一代测序应用和高通量SNP基因分型检测暴露了通常从单个蚊子个体获得的DNA量的局限性。全基因组扩增可以缓解这个问题，但它也会在基因组代表性方面产生偏差。在试图寻找提高DNA产量的替代DNA提取方案时，我们发现，将赛默飞世尔科技公司的组织裂解方案与Qiagen公司的DNA提取方案相结合，比仅使用Qiagen或赛默飞世尔科技公司的试剂盒方案能产生更多的DNA。我们尚未对提取方案的所有可能组合进行严格测试；我们也仅在蚊子样本上进行了测试。因此，应将我们的发现视为改进个人DNA提取方案的建议，而不是对市售产品的宣传。
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