聚合酶链反应产物特异性的阳性证实

Positive Identification of the Specificity of Polymerase Chain Reaction Product

以聚合酶链反应(PCR)法在mRNA水平检测T淋巴细胞受体α链可变区基因表达为例,介绍用^(32)P标记的人工合成寡核苷酸探针对PCR产物特异性作阳性证实的方法。该法以干琼脂糖凝胶作为支持物、相对较为简便和省财。用Ca探针以干凝胶作支持物的杂交结果,证实29个Vα基因的PCR扩增中物均为特异性的,放射自显影的带型与位置和溴乙锭染色所示完全吻合。

A method of positive identi-fication of the specificity of polymerasechain reaction(PCR) product using inter-nat oligonucleotide probe is introduced.The hybridization was done on the agaro-se gel which was dried after electropho-resis. Detection of the expression of T cellreceptor a chain variable (TCR Vα) geneson mRNA level was used as the experi-mental model. Twenty nine TCR Vα genesubfamilies could be distinguished clearlyin healthy human peripheral blood lym-phocytes by this method. Positive identifi-cation of PCR product on dried agarosegel by internal oligonucleotide probe is re-latively simple and less time consuming.