A DNA extraction protocol for improved DNA yield from individual mosquitoes [version 1; referees: 2 approved]

• Main Authors: Catelyn C. Nieman, Youki Yamasaki, Travis C. Collier, Yoosook Lee
• Format: Article
• Language: English
• Published: F1000 Research Ltd 2015-11-01
• Series: F1000Research
• Subjects: Animal Genetics; Control of Gene Expression
• Online Access: http://f1000research.com/articles/4-1314/v1

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.