Comparison of DNA extraction methods for PCR amplification of mitochondrial cytochrome c oxidase subunit II (COII) DNA from primate fecal samples

Christopher A. Whittier; Arun K. Dhar; Chip Stem; Jane Goodall; Acacia Alcivar-Warren

doi:10.1023/A:1008956921159

Comparison of DNA extraction methods for PCR amplification of mitochondrial cytochrome c oxidase subunit II (COII) DNA from primate fecal samples

Christopher A. Whittier, Arun K. Dhar, Chip Stem, Jane Goodall, Acacia Alcivar-Warren

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

16 Scopus citations

Abstract

Mitochondrial COII DNA was amplified by PCR from total DNA extracted from field collected primate fecal samples (n = 24) which had been stored without refrigeration for over 30 days. High molecular weight DNA total DNA was obtained from samples stored in 70% (v/v) ethanol, SDS lysis buffer (LB) and guanidine isothiocyanate buffer (GTB) than from samples stored in 10% formalin. Fecal DNA quality and COII amplification varied according to storage solution (formalin, ethanol, LB and GTB), extraction method (LB-based and GTB-based) and primate species (chimpanzee, baboon, human). It is recommended that fecal samples be collected in LB for DNA analysis. However, GTB-based protocols are suitable when total RNA is needed for epidemiological studies of viral diseases or gene expression analysis.

Original language	English (US)
Pages (from-to)	771-779
Number of pages	9
Journal	Biotechnology Techniques
Volume	13
Issue number	11
DOIs	https://doi.org/10.1023/A:1008956921159
State	Published - 1999
Externally published	Yes

Keywords

Baboon
COII
Chimpanzee
DNA
Fecal
Human

ASJC Scopus subject areas

Biochemistry
Applied Microbiology and Biotechnology

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10.1023/A:1008956921159

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title = "Comparison of DNA extraction methods for PCR amplification of mitochondrial cytochrome c oxidase subunit II (COII) DNA from primate fecal samples",

abstract = "Mitochondrial COII DNA was amplified by PCR from total DNA extracted from field collected primate fecal samples (n = 24) which had been stored without refrigeration for over 30 days. High molecular weight DNA total DNA was obtained from samples stored in 70% (v/v) ethanol, SDS lysis buffer (LB) and guanidine isothiocyanate buffer (GTB) than from samples stored in 10% formalin. Fecal DNA quality and COII amplification varied according to storage solution (formalin, ethanol, LB and GTB), extraction method (LB-based and GTB-based) and primate species (chimpanzee, baboon, human). It is recommended that fecal samples be collected in LB for DNA analysis. However, GTB-based protocols are suitable when total RNA is needed for epidemiological studies of viral diseases or gene expression analysis.",

keywords = "Baboon, COII, Chimpanzee, DNA, Fecal, Human",

author = "Whittier, {Christopher A.} and Dhar, {Arun K.} and Chip Stem and Jane Goodall and Acacia Alcivar-Warren",

note = "Funding Information: The authors would like to thank Drs R. Fister, D.A. Collins, F. Nutter and the field staff of the Gombe Stream Research Center and Tanzania National Parks for their assistance. Funding for this research was provided by NIH Training Grant T35DK07635 (C.W.), the Section of International Veterinary Medicine at TUSVM (C.W and C.S.), and generous grants from The Conservation, Food and Health Foundation and Geraldine R. Dodge Foundation Frontiers for Veterinary Medicine (C.W.) and the Curriculum Program at Tufts University School of Veterinary Medicine (A.W.).",

year = "1999",

doi = "10.1023/A:1008956921159",

language = "English (US)",

volume = "13",

pages = "771--779",

journal = "Biotechnology Techniques",

issn = "0951-208X",

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TY - JOUR

T1 - Comparison of DNA extraction methods for PCR amplification of mitochondrial cytochrome c oxidase subunit II (COII) DNA from primate fecal samples

AU - Whittier, Christopher A.

AU - Dhar, Arun K.

AU - Stem, Chip

AU - Goodall, Jane

AU - Alcivar-Warren, Acacia

N1 - Funding Information: The authors would like to thank Drs R. Fister, D.A. Collins, F. Nutter and the field staff of the Gombe Stream Research Center and Tanzania National Parks for their assistance. Funding for this research was provided by NIH Training Grant T35DK07635 (C.W.), the Section of International Veterinary Medicine at TUSVM (C.W and C.S.), and generous grants from The Conservation, Food and Health Foundation and Geraldine R. Dodge Foundation Frontiers for Veterinary Medicine (C.W.) and the Curriculum Program at Tufts University School of Veterinary Medicine (A.W.).

PY - 1999

Y1 - 1999

N2 - Mitochondrial COII DNA was amplified by PCR from total DNA extracted from field collected primate fecal samples (n = 24) which had been stored without refrigeration for over 30 days. High molecular weight DNA total DNA was obtained from samples stored in 70% (v/v) ethanol, SDS lysis buffer (LB) and guanidine isothiocyanate buffer (GTB) than from samples stored in 10% formalin. Fecal DNA quality and COII amplification varied according to storage solution (formalin, ethanol, LB and GTB), extraction method (LB-based and GTB-based) and primate species (chimpanzee, baboon, human). It is recommended that fecal samples be collected in LB for DNA analysis. However, GTB-based protocols are suitable when total RNA is needed for epidemiological studies of viral diseases or gene expression analysis.

AB - Mitochondrial COII DNA was amplified by PCR from total DNA extracted from field collected primate fecal samples (n = 24) which had been stored without refrigeration for over 30 days. High molecular weight DNA total DNA was obtained from samples stored in 70% (v/v) ethanol, SDS lysis buffer (LB) and guanidine isothiocyanate buffer (GTB) than from samples stored in 10% formalin. Fecal DNA quality and COII amplification varied according to storage solution (formalin, ethanol, LB and GTB), extraction method (LB-based and GTB-based) and primate species (chimpanzee, baboon, human). It is recommended that fecal samples be collected in LB for DNA analysis. However, GTB-based protocols are suitable when total RNA is needed for epidemiological studies of viral diseases or gene expression analysis.

KW - Baboon

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KW - Chimpanzee

KW - DNA

KW - Fecal

KW - Human

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Comparison of DNA extraction methods for PCR amplification of mitochondrial cytochrome c oxidase subunit II (COII) DNA from primate fecal samples

Abstract

Keywords

ASJC Scopus subject areas

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