【摘要】 目的:采用载体构建及荧光定量PCR的方法比较BCL2等位基因不同部位PCR引物在定量分析中的扩增效率,判断其用于不同等位基因转录活性分析的可靠性。方法:用PCR扩增包含定量PCR目的片段的序列,然后将扩增产物分别插入pGEM-TEasy载体中,构建TA3重组载体,再以TA3载体为模板,采用荧光定量PCR方法比较引物对之间扩增效率。结果:定量PCR结果显示引物对之间扩增效率无明显差异。结论:所检测的PCR引物对之间的扩增效率一致,可用于比较分析mbr+/mbr-Nalm-6杂合子细胞系中两个等位基因表达活性。更多还原

【Abstract】 Objective: To investigate the amplification efficiency of primers on different sites of BCL2 alleles in real-time PCR reactions and the reliability of these primers in analysis of transcription activity of different alleles with vector construction and real-time PCR. Methods: Fragments containing all amplicons from real-time PCR reactions were amplified by PCR and then inserted into the pGEM-T Easy vector to construct TA3 vector. The real-time PCR analysis was performed with TA3 plasmid as templates for testing the amplification efficiency of primers. Results: Real-time PCR showed no significant difference in the amplification efficiency for the of primers. Conclusion: Amplification efficiency of primers in real-time PCR reactions is identical. These primers can be used for quantity analysis of transcription activity of two BCL2 alleles in mbr+/mbr-Nalm-6 heterozygous cell line.更多还原