A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots

Víctor Otaño-Rivera, Amma Boakye, Nadja Grobe, Mohammed M. Almutairi, Shams Kursan, Lesan K. Mattis, Hayo Castrop, Susan Gurley, Khalid M. Elased, Gregory P. Boivin, Mauricio Di Fulvio

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Genotyping of genetically-engineered mice is necessary for the effective design of breeding strategies and identification of mutant mice. This process relies on the identification of DNA markers introduced into genomic sequences of mice, a task usually performed using the polymerase chain reaction (PCR). Clearly, the limiting step in genotyping is isolating pure genomic DNA. Isolation of mouse DNA for genotyping typically involves painful procedures such as tail snip, digit removal, or ear punch. Although the harvesting of hair has previously been proposed as a source of genomic DNA, there has been a perceived complication and reluctance to use this non-painful technique because of low DNA yields and fear of contamination. In this study we developed a simple, economic, and efficient strategy using Chelex® resins to purify genomic DNA from hair roots of mice which are suitable for genotyping. Upon comparison with standard DNA purification methods using a commercially available kit, we demonstrate that Chelex® efficiently and consistently purifies highquality DNA from hair roots, minimizing pain, shortening time and reducing costs associated with the determination of accurate genotypes. Therefore, the use of hair roots combined with Chelex® is a reliable and more humane alternative for DNA genotyping.

Original languageEnglish (US)
Pages (from-to)138-146
Number of pages9
JournalLaboratory Animals
Volume51
Issue number2
DOIs
StatePublished - Jan 1 2017
Externally publishedYes

Fingerprint

root hairs
Hair
Genotype
genomics
genotyping
genotype
DNA
mice
purification methods
fearfulness
Genetic Markers
shortenings
resins
Fear
hairs
Breeding
Ear
Tail
pain
ears

Keywords

  • 3Rs
  • Alternatives
  • DNA isolation
  • Rodents
  • Welfare

ASJC Scopus subject areas

  • Animal Science and Zoology
  • veterinary(all)

Cite this

Otaño-Rivera, V., Boakye, A., Grobe, N., Almutairi, M. M., Kursan, S., Mattis, L. K., ... Di Fulvio, M. (2017). A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots. Laboratory Animals, 51(2), 138-146. https://doi.org/10.1177/0023677216646088

A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots. / Otaño-Rivera, Víctor; Boakye, Amma; Grobe, Nadja; Almutairi, Mohammed M.; Kursan, Shams; Mattis, Lesan K.; Castrop, Hayo; Gurley, Susan; Elased, Khalid M.; Boivin, Gregory P.; Di Fulvio, Mauricio.

In: Laboratory Animals, Vol. 51, No. 2, 01.01.2017, p. 138-146.

Research output: Contribution to journalArticle

Otaño-Rivera, V, Boakye, A, Grobe, N, Almutairi, MM, Kursan, S, Mattis, LK, Castrop, H, Gurley, S, Elased, KM, Boivin, GP & Di Fulvio, M 2017, 'A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots', Laboratory Animals, vol. 51, no. 2, pp. 138-146. https://doi.org/10.1177/0023677216646088
Otaño-Rivera, Víctor ; Boakye, Amma ; Grobe, Nadja ; Almutairi, Mohammed M. ; Kursan, Shams ; Mattis, Lesan K. ; Castrop, Hayo ; Gurley, Susan ; Elased, Khalid M. ; Boivin, Gregory P. ; Di Fulvio, Mauricio. / A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots. In: Laboratory Animals. 2017 ; Vol. 51, No. 2. pp. 138-146.
@article{6e12d2010b484b31bb2c146ffbb0992d,
title = "A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots",
abstract = "Genotyping of genetically-engineered mice is necessary for the effective design of breeding strategies and identification of mutant mice. This process relies on the identification of DNA markers introduced into genomic sequences of mice, a task usually performed using the polymerase chain reaction (PCR). Clearly, the limiting step in genotyping is isolating pure genomic DNA. Isolation of mouse DNA for genotyping typically involves painful procedures such as tail snip, digit removal, or ear punch. Although the harvesting of hair has previously been proposed as a source of genomic DNA, there has been a perceived complication and reluctance to use this non-painful technique because of low DNA yields and fear of contamination. In this study we developed a simple, economic, and efficient strategy using Chelex{\circledR} resins to purify genomic DNA from hair roots of mice which are suitable for genotyping. Upon comparison with standard DNA purification methods using a commercially available kit, we demonstrate that Chelex{\circledR} efficiently and consistently purifies highquality DNA from hair roots, minimizing pain, shortening time and reducing costs associated with the determination of accurate genotypes. Therefore, the use of hair roots combined with Chelex{\circledR} is a reliable and more humane alternative for DNA genotyping.",
keywords = "3Rs, Alternatives, DNA isolation, Rodents, Welfare",
author = "V{\'i}ctor Ota{\~n}o-Rivera and Amma Boakye and Nadja Grobe and Almutairi, {Mohammed M.} and Shams Kursan and Mattis, {Lesan K.} and Hayo Castrop and Susan Gurley and Elased, {Khalid M.} and Boivin, {Gregory P.} and {Di Fulvio}, Mauricio",
year = "2017",
month = "1",
day = "1",
doi = "10.1177/0023677216646088",
language = "English (US)",
volume = "51",
pages = "138--146",
journal = "Laboratory Animals",
issn = "0023-6772",
publisher = "SAGE Publications Ltd",
number = "2",

}

TY - JOUR

T1 - A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots

AU - Otaño-Rivera, Víctor

AU - Boakye, Amma

AU - Grobe, Nadja

AU - Almutairi, Mohammed M.

AU - Kursan, Shams

AU - Mattis, Lesan K.

AU - Castrop, Hayo

AU - Gurley, Susan

AU - Elased, Khalid M.

AU - Boivin, Gregory P.

AU - Di Fulvio, Mauricio

PY - 2017/1/1

Y1 - 2017/1/1

N2 - Genotyping of genetically-engineered mice is necessary for the effective design of breeding strategies and identification of mutant mice. This process relies on the identification of DNA markers introduced into genomic sequences of mice, a task usually performed using the polymerase chain reaction (PCR). Clearly, the limiting step in genotyping is isolating pure genomic DNA. Isolation of mouse DNA for genotyping typically involves painful procedures such as tail snip, digit removal, or ear punch. Although the harvesting of hair has previously been proposed as a source of genomic DNA, there has been a perceived complication and reluctance to use this non-painful technique because of low DNA yields and fear of contamination. In this study we developed a simple, economic, and efficient strategy using Chelex® resins to purify genomic DNA from hair roots of mice which are suitable for genotyping. Upon comparison with standard DNA purification methods using a commercially available kit, we demonstrate that Chelex® efficiently and consistently purifies highquality DNA from hair roots, minimizing pain, shortening time and reducing costs associated with the determination of accurate genotypes. Therefore, the use of hair roots combined with Chelex® is a reliable and more humane alternative for DNA genotyping.

AB - Genotyping of genetically-engineered mice is necessary for the effective design of breeding strategies and identification of mutant mice. This process relies on the identification of DNA markers introduced into genomic sequences of mice, a task usually performed using the polymerase chain reaction (PCR). Clearly, the limiting step in genotyping is isolating pure genomic DNA. Isolation of mouse DNA for genotyping typically involves painful procedures such as tail snip, digit removal, or ear punch. Although the harvesting of hair has previously been proposed as a source of genomic DNA, there has been a perceived complication and reluctance to use this non-painful technique because of low DNA yields and fear of contamination. In this study we developed a simple, economic, and efficient strategy using Chelex® resins to purify genomic DNA from hair roots of mice which are suitable for genotyping. Upon comparison with standard DNA purification methods using a commercially available kit, we demonstrate that Chelex® efficiently and consistently purifies highquality DNA from hair roots, minimizing pain, shortening time and reducing costs associated with the determination of accurate genotypes. Therefore, the use of hair roots combined with Chelex® is a reliable and more humane alternative for DNA genotyping.

KW - 3Rs

KW - Alternatives

KW - DNA isolation

KW - Rodents

KW - Welfare

UR - http://www.scopus.com/inward/record.url?scp=85018370723&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85018370723&partnerID=8YFLogxK

U2 - 10.1177/0023677216646088

DO - 10.1177/0023677216646088

M3 - Article

VL - 51

SP - 138

EP - 146

JO - Laboratory Animals

JF - Laboratory Animals

SN - 0023-6772

IS - 2

ER -