Plucked hair samples as a source of DNA: reliability of dinucleotide microsatellite genotyping

被引:191
作者
Goossens, B [1 ]
Waits, LP [1 ]
Taberlet, P [1 ]
机构
[1] Univ Grenoble 1, Lab Biol Populat Altitude, CNRS, UMR 5553, F-38041 Grenoble 9, France
关键词
genetic typing; hair; Marmota marmota; multiple-tubes approach; noninvasive genetic sampling; polymerase chain reaction;
D O I
10.1046/j.1365-294x.1998.00407.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
To test whether plucked hairs are a reliable source of DNA for genotyping microsatellite loci, we carried out experiments using one, three, or 10 hairs per extract for 50 alpine marmots. For each extract, seven independent genotypings were performed for the same locus (multiple-tubes approach). Two types of genotyping errors were recorded: a false homozygote defined as the detection of only one allele of a true heterozygote, and a false allele defined as a PCR-generated allele that was not one of the alleles of the true genotype. Using DNA extracted from one, three, or 10 hairs, the overall error rate was 14.00%, 4.86%, and 0.29%, respectively. Based on our results, we conclude that 10 hairs should be used to obtain consistently reliable genotypings using the single-tube approach, and that a single plucked hair could represent a reliable source of DNA if the multiple-tubes approach is used. For future studies of dinucleotide repeat diversity using DNA extracted from one to three shed or plucked hairs, we strongly recommend initiating,an appropriate pilot study to quantify the error rate and to determine the reliability of the single-tube approach.
引用
收藏
页码:1237 / 1241
页数:5
相关论文
共 15 条
  • [1] CALLEN DF, 1993, AM J HUM GENET, V52, P922
  • [2] EDWARDS A, 1991, AM J HUM GENET, V49, P746
  • [3] Experimental limits of PCR analysis of (CA)(n) repeat alterations
    Foucault, F
    Praz, F
    Jaulin, C
    AmorGueret, M
    [J]. TRENDS IN GENETICS, 1996, 12 (11) : 450 - 452
  • [4] Microsatellite scoring errors associated with noninvasive genotyping based on nuclear DNA amplified from shed hair
    Gagneux, P
    Boesch, C
    Woodruff, DS
    [J]. MOLECULAR ECOLOGY, 1997, 6 (09) : 861 - 868
  • [5] AMPLIFICATION OF HYPERVARIABLE SIMPLE SEQUENCE REPEATS (MICROSATELLITES) FROM EXCREMENTAL DNA OF WILD LIVING BONOBOS (PAN-PANISCUS)
    GERLOFF, U
    SCHLOTTERER, C
    RASSMANN, K
    RAMBOLD, I
    HOHMANN, G
    FRUTH, B
    TAUTZ, D
    [J]. MOLECULAR ECOLOGY, 1995, 4 (04) : 515 - 518
  • [6] Goossens B, 1996, CR ACAD SCI III-VIE, V319, P241
  • [7] KLINKICHT M, 1993, THESIS U MUNICH
  • [8] Morin Phillip A., 1996, P298
  • [9] NAVIDI W, 1992, AM J HUM GENET, V50, P347
  • [10] How many cycles does a PCR need? Determinations of cycle numbers depending on the number of targets and the reaction efficiency factor
    Rameckers, J
    Hummel, S
    Herrmann, B
    [J]. NATURWISSENSCHAFTEN, 1997, 84 (06) : 259 - 262