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目的 以并行检测单核苷酸多态性 (SNP)为基础 ,研制对牛个体进行可靠鉴定的遗传学条形码生物芯片。方法 将生物素化等位基因特异性寡核苷酸固定于生物芯片上长链烷烃硫醇自组装单层已连接链霉亲和素 (SA)的部位。多重PCR法扩增待杂交并于芯片上微型测序的靶DNA。结果 经过微型测序的酶促延伸反应 ,能可靠地从待测基因检出所有C/T碱基替换。结论 我们的SNP芯片系统能用于检测任何基因的C/T点突变
Objective To develop a genetic barcode biochip for the reliable identification of cattle individuals based on the parallel detection of single nucleotide polymorphisms (SNPs). Methods The biotinylated allele-specific oligonucleotides were immobilized on the biochip where the long-chain alkanethiol self-assembled monolayers were linked to streptavidin (SA). Multiplex PCR amplification of target DNA to be hybridized and micro-sequenced on-chip. Results Microdissected enzymatic extension allows reliable detection of all C / T base substitutions from the gene to be tested. Conclusion Our SNP chip system can be used to detect C / T point mutations in any gene