Back to Search Start Over

A DNA extraction protocol for improved DNA yield from individual mosquitoes [version 1; referees: 3 approved]

Authors :
Catelyn C. Nieman
Youki Yamasaki
Travis C. Collier
Yoosook Lee
Author Affiliations :
<relatesTo>1</relatesTo>Vector Genetics Laboratory, Department of Pathology, Microbiology, and Immunology, University of California, Davis, CA, USA
Source :
F1000Research. 4:1314
Publication Year :
2015
Publisher :
London, UK: F1000 Research Limited, 2015.

Abstract

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.

Details

ISSN :
20461402
Volume :
4
Database :
F1000Research
Journal :
F1000Research
Notes :
[version 1; referees: 3 approved]
Publication Type :
Academic Journal
Accession number :
edsfor.10.12688.f1000research.7413.1
Document Type :
method-article
Full Text :
https://doi.org/10.12688/f1000research.7413.1