摘要
通过设计通用荧光PCR引物并结合DNA测序系统建立了小鼠的多重STR分型方案。实验针对小鼠基因组设计了两对不同的通用引物序列,标记了FAM荧光的通用序列和"加尾"的位点特异性引物共同用于小鼠的多重PCR的STR基因分型。本研究优化了通用引物和特异性引物间的比例,优化了多重STR-PCR的反应条件,并最终利用该技术方案实现了五重STR分型。实验验证了该方案在多重STR分型中的可行性。与传统的荧光检测PCR产物方案相比,应用通用方案完成多重PCR反应大大节省了实验时间与经费。
This study established a multiplex STR genotyping system through universal fluorescent primers in combination with sequencing. For each panel of multiplex STR genotyping amplification, two sets of primers were used, namely universal primers labeled with FAM and specific primers with an added 5＇ tail. We optimized the proportion of universal and specific primers and reaction of multiplex PCR was achieved. By using this method, five STR marker loci genotyping were successfully conducted within a single reaction. The optimized universal fluorescent primers amplification genotyping system can be used in multiplex STR genotyping, which allows reliable, effective and low-cost genotyping compared with regular microsatellite fluorescent detection assays.
出处
《动物学杂志》
CAS
CSCD
北大核心
2008年第3期75-80,共6页
Chinese Journal of Zoology
基金
国家自然科学基金项目(No.30771191)
关键词
通用引物
STR
多重PCR
基因分型
Universal primers
STR
Multiplex polymerase chain reaction
Genotyping
