To detect multiplex single nucleotide polymorphisms(SNPs)simultaneously,a new method was established by combining ALM-ASA with microfabricated CE-chip.Taking the CYP2D6 gene as an example,six SNPs,100C>T(P34S),1707...To detect multiplex single nucleotide polymorphisms(SNPs)simultaneously,a new method was established by combining ALM-ASA with microfabricated CE-chip.Taking the CYP2D6 gene as an example,six SNPs,100C>T(P34S),1707T>del(frameshift),1758G>T(stop codon),2470T>C,2549A>del(frameshift)and 2613AGA>del(K281del),were typed by four steps consisting of preamplification,digestion and ligation,allele-specific amplification,and amplicon separation by chip-CE.The genotyping results of 20 different genome samples by 6-plexed ALM-ASA were completely consistent with those obtained by polymerase chain reaction-restriction fragment length polymorphism analysis(PCR-RFLP),indicating that the multiplex approach established in this study was accurate and inexpensive.As the small reagent consumption by CE-chip device,a low cost for SNP typing was achieved together with the multiplex PCR technology proposed in this report.Neither modification of microchip channels nor clean-up process of PCR products was required;this greatly shortens the whole time for SNP typing.展开更多
文摘To detect multiplex single nucleotide polymorphisms(SNPs)simultaneously,a new method was established by combining ALM-ASA with microfabricated CE-chip.Taking the CYP2D6 gene as an example,six SNPs,100C>T(P34S),1707T>del(frameshift),1758G>T(stop codon),2470T>C,2549A>del(frameshift)and 2613AGA>del(K281del),were typed by four steps consisting of preamplification,digestion and ligation,allele-specific amplification,and amplicon separation by chip-CE.The genotyping results of 20 different genome samples by 6-plexed ALM-ASA were completely consistent with those obtained by polymerase chain reaction-restriction fragment length polymorphism analysis(PCR-RFLP),indicating that the multiplex approach established in this study was accurate and inexpensive.As the small reagent consumption by CE-chip device,a low cost for SNP typing was achieved together with the multiplex PCR technology proposed in this report.Neither modification of microchip channels nor clean-up process of PCR products was required;this greatly shortens the whole time for SNP typing.
