生物素掺入PCR产物的反向杂交技术分析HLA基因型

HLA-DPB Genotyped by Reverse Slot Blots Hybridization with Biotin-incorporated PCR Product

本文介绍了与常规生物素或荧光标记引物不同，以及与同位素掺入ＰＣＲ产物不同的反向杂交技术，研究了生物素最佳掺入条件，测出加尾探针联结到尼龙膜上所需紫外光的强度，比较了生物素５＇端标记引物与生物素掺入ＰＣＲ产物的显色灵敏度，并根据实验测得的最佳条件分析了３个标准细胞株ＨＬＡ－ＤＰＢ_１基因型。

A new technique of reverse slot blots hybridization with biotin incorporated PCR product, which is different from that produced by biotin or fluoresent labeled primer, has been applied to HLA-DPB genotyping of 3 B-lymphocyte lines (LUY, E4181324,EHM). We found the best condition for incorporating biotin-11-dUTP into PCR product, the suitable UV dosage for binding tailed oligonucleotides probes to nylon fliter and finally, compared colorimetric sensitivity of PCR product by biotin-labeled primer with that by biotin-incorporation.