非同位素标记探针杂交分析白细胞端粒DNA长度

目的建立用非同位素标记探针杂交测定端粒DNA长度的方法,并探讨其法医学的应用价值。方法酚/氯仿抽提基因组DNA,限制性内切酶消化,0.8％琼脂糖凝胶电泳,Southern印迹,化学发光法检测端粒DNA谱带,与标准分子量DNA比较,积分光密度扫描计算端粒DNA平均长度。结果所测样本获得了较好的低背景杂交谱带,测得31~35岁端粒。DNA的平均长度为11.71kb,51~55岁平均为11.04 kb。结论用上述建立的方法初步显示年龄与端粒长度之间有一定的相关性,为法医学年龄的推断开辟了一个新的研究领域。

Telomere length measurement of human leukocyte in peripheral blood by non-radio -isotopic hybridization

Objective To measure telomere length by hybridization with non - radio - isotopic - labeled probes and study its application in forensic medicine. Methods Genomic DNA from 60 donors (aged 31 ~ 35 years - old,51 ~ 55years - old) was digested by Hinf I /Rsa I and hybridized by digoxin - labeled probes, then chemo - luminescent substrates showed the blot, and the mean TRF length was calculated by comparing the signals relative to a molecular weight standard.Results The TRF of 31 ~ 35 years - old persons was 11.71kb and 11.04kb between 51 and 55 years - old persons.Conclusion A digoxin - labeled and low background method for determining telomere length was established, which suggested that the relationship might be existed in age and telomere length.