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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。
Using polymerase chain reaction (PCR) to detect the gene expression of the variable region of α-chain of T lymphocyte receptor at the mRNA level as an example, the PCR product specificities of ~ (32) P labeled synthetic oligonucleotide probes For positive confirmation of the method. The method of dry agarose gel as a supporter, relatively simple and save money. The results of the hybridization of the Ca probe with the xerogel as a support confirmed that all of the 29 Vα genes were specific for PCR amplification and that the autoradiogram pattern exactly matches the position and ethidium bromide staining.