荧光定量PCR测定端粒长度

目的 应用实时荧光定量聚合酶链式反应(Q-PCR)方法测定端粒长度.方法 选取9种人类细胞株,提取基因组DNA,采用Q-PCR方法测定相对T/S比率,DNA印迹法测定末端限制性片段(TRF)长度,进行二者之间的相关性分析.结果 定量PCR测定端粒长度相对T/S比率为0.68±0.57,DNA印迹法测量平均TRF值为8.57±2.34,两种方法测定结果的相关性分析R2=0.7807(P＜0.01).结论 采用荧光定量PCR方法测量端粒长度具有重复性好、省时、简便、可靠的特点,可高通量处理大量样品.

Telomere Measurement by Quantitative PCR

Objective To determine relative telomere length by quantitative PCR.Methods Genomic DNA was extracted from 9 types of human cells.Relative T/S ratios measured by quantitative PCR were compared with relative mean terminal restriction fragment(TRF)lengths in the same samples measured by traditional Southern blot methods.Results Relative R/S ratio measured by Q-PCR was 0.68±0.57,and relative mean TRF length was 8.57±2.34.By linear regression analysis,the correlation coefficient,R~2,for the relationship of T/S ratio to TRF length,was 0.7807(P＜0.001).Conclusion We have demonstrated the measurement of relative average telomere lengths by quantitative PCR,using a carefully designed pair of oligonucleotide primers.The assay is simple,rapid and reproducible,thus reliable for a high throughput of samples.It is recommendable to be used in study of telomere biology and genetic epidemiology of cancer and aging related diseases.