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A DNA extraction protocol for improved DNA yield from individual mosquitoes
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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
F1000Research
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4743141/ https://www.ncbi.nlm.nih.gov/pubmed/26937269 http://dx.doi.org/10.12688/f1000research.7413.1 |
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author | Nieman, Catelyn C. Yamasaki, Youki Collier, Travis C. Lee, Yoosook |
author_facet | Nieman, Catelyn C. Yamasaki, Youki Collier, Travis C. Lee, Yoosook |
author_sort | Nieman, Catelyn C. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4743141 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | F1000Research |
record_format | MEDLINE/PubMed |
spelling | pubmed-47431412016-03-01 A DNA extraction protocol for improved DNA yield from individual mosquitoes Nieman, Catelyn C. Yamasaki, Youki Collier, Travis C. Lee, Yoosook F1000Res Method Article 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. F1000Research 2015-11-20 /pmc/articles/PMC4743141/ /pubmed/26937269 http://dx.doi.org/10.12688/f1000research.7413.1 Text en Copyright: © 2015 Nieman CC et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Method Article Nieman, Catelyn C. Yamasaki, Youki Collier, Travis C. Lee, Yoosook A DNA extraction protocol for improved DNA yield from individual mosquitoes |
title | A DNA extraction protocol for improved DNA yield from individual mosquitoes |
title_full | A DNA extraction protocol for improved DNA yield from individual mosquitoes |
title_fullStr | A DNA extraction protocol for improved DNA yield from individual mosquitoes |
title_full_unstemmed | A DNA extraction protocol for improved DNA yield from individual mosquitoes |
title_short | A DNA extraction protocol for improved DNA yield from individual mosquitoes |
title_sort | dna extraction protocol for improved dna yield from individual mosquitoes |
topic | Method Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4743141/ https://www.ncbi.nlm.nih.gov/pubmed/26937269 http://dx.doi.org/10.12688/f1000research.7413.1 |
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