Electrophoretic detection of single-nucleotide polymorphisms

被引:21
作者
See, D [1 ]
Kanazin, V [1 ]
Talbert, H [1 ]
Blake, T [1 ]
机构
[1] Montana State Univ, Dept Plant Sci, Bozeman, MT 59717 USA
关键词
D O I
10.2144/00284st07
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Single-nucleotide polymorphisms (SNPs) represent the most prevalent class of genetic markers available for linkage disequilibrium or cladistic analyses. PCR primers may be labeled with fluorescent dyes and used to rapidly and accurately differentiate among alleles that are defined by a single-nucleotide differences. Here, we describe the primer-mediated detection of SNPs based on primer mismatch during allele-specific amplification of preamplified target sequences. Primers are labeled with different fluors at their 5' nucleotides, with their 3' termini at the transition mutation that defines allelic variation at the target locus. Each primer perfectly matches one of the two available alleles Sor each locus. Electrophoretic detection pel-mits characterization of the product both by size and fluor. This report demonstrates some of the capabilities of this assay, including heterozygote determination and multiplexed analysis.
引用
收藏
页码:710 / +
页数:5
相关论文
共 18 条
[1]   STS-PCR markers appropriate for wheat-barley introgression [J].
Blake, TK ;
Kadyrzhanova, D ;
Shepherd, KW ;
Islam, AKMR ;
Langridge, PL ;
McDonald, CL ;
Erpelding, J ;
Larson, S ;
Blake, NK ;
Talbert, LE .
THEORETICAL AND APPLIED GENETICS, 1996, 93 (5-6) :826-832
[2]   Template-directed dye-terminator incorporation (TDI) assay: A homogeneous DNA diagnostic method based on fluorescence resonance energy transfer [J].
Chen, XN ;
Kwok, PY .
NUCLEIC ACIDS RESEARCH, 1997, 25 (02) :347-353
[3]  
Dellaporta S.L., 1983, Plant Molecular Biology Reporter, V1, P19, DOI DOI 10.1007/BF02712670
[4]   Single nucleotide polymorphic discrimination by an electronic dot blot assay on semiconductor microchips [J].
Gilles, PN ;
Wu, DJ ;
Foster, CB ;
Dillon, PJ ;
Chanock, SJ .
NATURE BIOTECHNOLOGY, 1999, 17 (04) :365-370
[5]   Single-nucleotide polymorphism identification assays using a thermostable DNA polymerase and delayed extraction MALDI-TOF mass spectrometry [J].
Haff, LA ;
Smirnov, IP .
GENOME RESEARCH, 1997, 7 (04) :378-388
[6]  
HAYES PM, 1994, THEOR APPL GENET, V87, P397
[7]   A MOLECULAR, ISOZYME AND MORPHOLOGICAL MAP OF THE BARLEY (HORDEUM-VULGARE) GENOME [J].
KLEINHOFS, A ;
KILIAN, A ;
MAROOF, MAS ;
BIYASHEV, RM ;
HAYES, P ;
CHEN, FQ ;
LAPITAN, N ;
FENWICK, A ;
BLAKE, TK ;
KANAZIN, V ;
ANANIEV, E ;
DAHLEEN, L ;
KUDRNA, D ;
BOLLINGER, J ;
KNAPP, SJ ;
LIU, B ;
SORRELLS, M ;
HEUN, M ;
FRANCKOWIAK, JD ;
HOFFMAN, D ;
SKADSEN, R ;
STEFFENSON, BJ .
THEORETICAL AND APPLIED GENETICS, 1993, 86 (06) :705-712
[8]   Reading bits of genetic information: Methods for single-nucleotide polymorphism analysis [J].
Landegren, U ;
Nilsson, M ;
Kwok, PY .
GENOME RESEARCH, 1998, 8 (08) :769-776
[9]  
Larson SR, 1996, THEOR APPL GENET, V93, P618, DOI 10.1007/BF00417957
[10]   Overlapping PCR for bidirectional PCR amplification of specific alleles: A rapid one-tube method for simultaneously differentiating homozygotes and heterozygotes [J].
Liu, Q ;
Thorland, EC ;
Heit, JA ;
Sommer, SS .
GENOME RESEARCH, 1997, 7 (04) :389-398