论文部分内容阅读
本研究运用异硫氰酸胍-酚-乙醇法提取人睾丸组织中的RNA,并用高效Oligo(dt)cellulose从中纯化mRNA,以此为模板,在AMV反转录酶作用下,制备互补的DNA(ss-cDNA),RNaseH降解模板mRNA,经DNA多聚酶I作用以ss-cDNA为模板合成第二链(ds-cDNA)。用T_4DNA多聚酶制备cDNA的平末端,在T_4DNA连接酶的
In this study, guanidine isothiocyanate-phenol-ethanol method was used to extract RNA from human testicular tissue and purified from the mRNA by high-performance Oligo (dt) cellulose as a template to prepare complementary DNA under the action of AMV reverse transcriptase (ss-cDNA), RNaseH degraded template mRNA, and the second strand (ds-cDNA) was synthesized by DNA polymerase I using ss-cDNA as a template. The T_4 DNA polymerase was used to prepare the blunt end of the cDNA in T_4 DNA ligase