摘要
背景:以往聚合酶链反应产物检测多采用EB染色、银染色等技术,由于肉眼观察容易产生误差,尤其对片段长度差距较小、谱带位置相距较近的电泳图谱分析更加困难。目的:通过ABI310遗传分析仪检测多重聚合酶链反应扩增短串联重复序列产物,评价其数据分析的准确性。设计、时间及地点:2003-09/2004-01在四川农业大学分子遗传实验室完成。材料:凉山半细毛羊20只,取耳组织。方法:采用多重重聚合酶链反应扩增凉山半细毛羊基因组上的Y染色体上SRY,MCM158,MAF45和X染色体上MILVET09,AE25序列后,进行ABI310遗传分析仪检测。主要观察指标:5个基因座的基因型和片断大小。结果:通过ABI310遗传分析仪检测准确获得5个基因座的全自动分型结果和片断大小。结论:ABI310遗传分析仪检测多重聚合酶链反应扩增短串联重复序列产物,得到的数据准确可靠。
BACKGROUND： EB staining and silver staining are commonly used in previous detection of PCR product. However, macroscopic observation always leads to errors, especially for analyses of electrophoretogram of similar fragment length and close gaps. OBJECTIVE： To evaluate the accuracy of data analyses process by ABI 310 Genetic Analyzer in multiple polymerase chain reaction and short tandem repeat （PCR-STR）. DESIGN, TIME AND SETTING： The experiment was performed at the Molecular Genetics Laboratory of Sichuan Agricultural University between September 2003 and January 2004. MATERIALS： Twenty Liangshan semi-fine wool sheep were selected and ear tissues were harvested. METHODS： The technique of multiple PCR was used to amplify the microsatellite markers MILVET09 and AE25 on X chromosome and MCM158 and MAF45 on Y chromosome and SRY gene of the sheep, then the amplified products were measured by ABI 310 Genetic Analyzer. MAIN OUTCOME MEASURES： Genotypes and gene fragment sizes of five loci. RESULTS： The results of genotyping and fragment sizes of five loci were obtained accurately by ABI 310 Genetic Analyzer. CONCLUSION： ABI 310 Genetic Analyzer could accurately detect the amplified products by multiple PCR.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第35期6859-6862,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
