[A method of haplotype analysis for multiple single-nucleotide polymorphisms].
Haplotypes from multiple single nucleotide polymorphisms(SNPs) spaced in longer DNA were constructed by multi-step PCR and DNA sequencing methods. Two allele-specific primers were synthesized and used for long DNA fragments (approximately 10 kb) amplification from human genome DNA. Fragments within these long DNA fragments were amplified by using these PCR products as templates in the second round PCR. The second round PCR products were subsequently sequenced. Haplotype construction was performed based on the character of nucleotides at the 3'-end of the allele-specific primers and the sequencing results from the second round PCR products. The DNA fragment (approximately 16 kb) from promoter to exon 4 of lipoprotein lipase (LPL) gene was amplified by allele-specific primers,and the DNA fragments including exon 2 or exon 3 of LPL gene were amplified and sequenced. A SNP of +13,557G-->A within intron 2 was identified. Four haplotypes including -421 G/ +13557G/ + 15222A, - 421A/ +13557G/ +15222A, -421G/ +13557G/ + 15222G, - 421G/+ 13557A/ +15222A were detected. The method is effective and relatively simple for construction of haplotypes from multiple single nucleotide polymorphisms.
Su
Zhi-Guang
ZG
Department of Medical Genetics, West China Hospital, Sichuan University, Chengdu 610041, China. zhiguang_su@hotmail.com
Zhang
Si-Zhong
SZ
Xiao
Cui-Ying
CY
Tong
Yu
Y
D000483:Alleles; D001483:Base Sequence; D003327:Coronary Disease / Q000235:genetics; D005091:Exons; D005260:Female; D006239:Haplotypes / Q000235:genetics*; D006801:Humans; D007438:Introns; D008071:Lipoprotein Lipase / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA
D008071:Lipoprotein Lipase