A DNA extraction protocol for improved DNA yield from individual mosquitoes

Yoosook Lee, Catelyn C. Nieman, Youki Yamasaki, Travis C. Collier

Research output: Contribution to journalArticle

9 Citations (Scopus)

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.

Original languageEnglish (US)
Article number1314
JournalF1000Research
Volume4
DOIs
StatePublished - Jan 1 2015

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Culicidae
DNA
Assays
Genes
Genome
Technology
Single Nucleotide Polymorphism
Amplification
Throughput
Tissue
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Medicine(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

A DNA extraction protocol for improved DNA yield from individual mosquitoes. / Lee, Yoosook; Nieman, Catelyn C.; Yamasaki, Youki; Collier, Travis C.

In: F1000Research, Vol. 4, 1314, 01.01.2015.

Research output: Contribution to journalArticle

Lee, Yoosook ; Nieman, Catelyn C. ; Yamasaki, Youki ; Collier, Travis C. / A DNA extraction protocol for improved DNA yield from individual mosquitoes. In: F1000Research. 2015 ; Vol. 4.
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