新型TaqMan-MB荧光探针测定古DNA模板含量

New type taqman-MB of fluorogenic probe for ancient DNA template detection

目的:设计荧光检测探针,对古DNA样本中的线粒体DNA片段进行定量分析,并对其检测效果进行评价。方法:利用分子信标的茎环结构和TaqMan的降解作用设计一种新型的TaqMan-Molecular Beacon荧光检测探针,与实时PCR仪联用,对古DNA模板进行定量检测。结果:5个古代样本中3个测得拷贝数,1个无法观察到原始模板量,1个样本无原始模板。结论:TaqMan-MB荧光探针集中了分子信标和TaqMan的技术优势,对古DNA模板具有很好的定量检测效果。

Objective To develop a fluorogenic probe for measuring the concentration of DNA extracted from ancient samples, and evaluate its detection effects . Methods A novel fluorogenic probe, TaqMan-Molecular Beacon （TaqMan MB）, which combined the hairpin structure of molecular beacons and the cleavage mechanism of TaqMan probes, was developed. Combined with a real-time PCR instrument, TagMan-MB was used for quantification of templates of ancient DNA. Results In all five ancient samples, three samples were gotten the copy numbers, one sample had no template, and one sample cannot be detected. Conclusion TaqMan-MB has much better detection effects than TaqMan or molecular beacons alone, and presents a promising approach for quantification of target genes.