Evaluation of a reliable and cost-effective method of DNA isolation for mouse genotyping

Justin R. Sysol; Carrie Kempf; Michelle N. Helton; Yangbasai Dong; Daocheng Zhu; Haiying Sun; Joe G.N. Garcia; Roberto F. Machado; Jiwang Chen

doi:10.1007/s10529-012-1113-2

Evaluation of a reliable and cost-effective method of DNA isolation for mouse genotyping

Justin R. Sysol, Carrie Kempf, Michelle N. Helton, Yangbasai Dong, Daocheng Zhu, Haiying Sun, Joe G.N. Garcia, Roberto F. Machado, Jiwang Chen

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Genotyping is commonly used to define specific gene alterations or the presence of transgenes in mice. This procedure is typically done using DNA isolated from mouse tail tissue. Although there are commercially available kits for tail DNA isolation, they can be time consuming and costly for routine genotyping. In this study, we describe a rapid, "crude" DNA isolation method using mouse tail tissue and compare it to a frequently used, commercially available kit in the genotyping of over 1,000 total mice from 8 genetic lines. Our genotyping results were obtained faster and less expensively but with the same success rate (Crude method: 97. 7 %, Kit method: 98. 4 %). To our knowledge, this is the first systematic study to compare the reliability of this crude DNA isolation method for mouse genotyping compared to a commercially available kit.

Original language	English (US)
Pages (from-to)	509-514
Number of pages	6
Journal	Biotechnology Letters
Volume	35
Issue number	4
DOIs	https://doi.org/10.1007/s10529-012-1113-2
State	Published - Apr 2013
Externally published	Yes

Keywords

Genomic DNA isolation
Knockout mice
Mouse genotyping
PCR
Tail biopsy

ASJC Scopus subject areas

Biotechnology
Bioengineering
Applied Microbiology and Biotechnology

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10.1007/s10529-012-1113-2

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title = "Evaluation of a reliable and cost-effective method of DNA isolation for mouse genotyping",

abstract = "Genotyping is commonly used to define specific gene alterations or the presence of transgenes in mice. This procedure is typically done using DNA isolated from mouse tail tissue. Although there are commercially available kits for tail DNA isolation, they can be time consuming and costly for routine genotyping. In this study, we describe a rapid, {"}crude{"} DNA isolation method using mouse tail tissue and compare it to a frequently used, commercially available kit in the genotyping of over 1,000 total mice from 8 genetic lines. Our genotyping results were obtained faster and less expensively but with the same success rate (Crude method: 97. 7 %, Kit method: 98. 4 %). To our knowledge, this is the first systematic study to compare the reliability of this crude DNA isolation method for mouse genotyping compared to a commercially available kit.",

keywords = "Genomic DNA isolation, Knockout mice, Mouse genotyping, PCR, Tail biopsy",

author = "Sysol, {Justin R.} and Carrie Kempf and Helton, {Michelle N.} and Yangbasai Dong and Daocheng Zhu and Haiying Sun and Garcia, {Joe G.N.} and Machado, {Roberto F.} and Jiwang Chen",

note = "Funding Information: Acknowledgments This study was supported by the North-western Faculty Foundation Dixon Innovation Award (North-western University, Chicago, IL) and the Pfizer ASPIRE (Advancing Science through Pfizer—Investigator Research Exchange) Award (Pfizer Inc.), both awarded to Dr. Jiwang Chen. We thank Dr. Youyang Zhao at the University of Illinois at Chicago for generously providing the eNOS knockout mice and primers used in this study.",

year = "2013",

month = apr,

doi = "10.1007/s10529-012-1113-2",

language = "English (US)",

volume = "35",

pages = "509--514",

journal = "Biotechnology Letters",

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AU - Sysol, Justin R.

AU - Kempf, Carrie

AU - Helton, Michelle N.

AU - Dong, Yangbasai

AU - Zhu, Daocheng

AU - Sun, Haiying

AU - Garcia, Joe G.N.

AU - Machado, Roberto F.

AU - Chen, Jiwang

N1 - Funding Information: Acknowledgments This study was supported by the North-western Faculty Foundation Dixon Innovation Award (North-western University, Chicago, IL) and the Pfizer ASPIRE (Advancing Science through Pfizer—Investigator Research Exchange) Award (Pfizer Inc.), both awarded to Dr. Jiwang Chen. We thank Dr. Youyang Zhao at the University of Illinois at Chicago for generously providing the eNOS knockout mice and primers used in this study.

PY - 2013/4

Y1 - 2013/4

N2 - Genotyping is commonly used to define specific gene alterations or the presence of transgenes in mice. This procedure is typically done using DNA isolated from mouse tail tissue. Although there are commercially available kits for tail DNA isolation, they can be time consuming and costly for routine genotyping. In this study, we describe a rapid, "crude" DNA isolation method using mouse tail tissue and compare it to a frequently used, commercially available kit in the genotyping of over 1,000 total mice from 8 genetic lines. Our genotyping results were obtained faster and less expensively but with the same success rate (Crude method: 97. 7 %, Kit method: 98. 4 %). To our knowledge, this is the first systematic study to compare the reliability of this crude DNA isolation method for mouse genotyping compared to a commercially available kit.

AB - Genotyping is commonly used to define specific gene alterations or the presence of transgenes in mice. This procedure is typically done using DNA isolated from mouse tail tissue. Although there are commercially available kits for tail DNA isolation, they can be time consuming and costly for routine genotyping. In this study, we describe a rapid, "crude" DNA isolation method using mouse tail tissue and compare it to a frequently used, commercially available kit in the genotyping of over 1,000 total mice from 8 genetic lines. Our genotyping results were obtained faster and less expensively but with the same success rate (Crude method: 97. 7 %, Kit method: 98. 4 %). To our knowledge, this is the first systematic study to compare the reliability of this crude DNA isolation method for mouse genotyping compared to a commercially available kit.

KW - Genomic DNA isolation

KW - Knockout mice

KW - Mouse genotyping

KW - PCR

KW - Tail biopsy

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Evaluation of a reliable and cost-effective method of DNA isolation for mouse genotyping

Abstract

Keywords

ASJC Scopus subject areas

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