【摘要】 以多条末端互补的寡核苷酸链为模板,用PCR技术能够一次扩增出目的基因片段.将预合成的350bp目的基因片段分为6条寡聚核苷酸链(S1～S6),每相邻两条寡聚核苷酸链的末端有18个碱基的互补区.先将S1S2,S3S4,S5S6分别用Taq酶延伸补平形成3条双链DNA;再将S1S2,S3S4,S5S6三条双链DNA以3∶1∶3的比例混合,用预先设计的含有多克隆位点的引物进行PCR扩增,一次性扩增出完整的350bp目的基因片段.更多还原

【Abstract】 An artifical gene fragments about 350 bases were quickly synthesized by PCR technology without any mistakes revealed by DNA sequencing. Initially, six oligos (S1～S6) were prepared, all about 80 bases in length, which have 18 bp complementary sequences to each other at their ends. Three short double strand DNAs (ds DNAs) were generated after annealing three pairs of oligos using Taq DNA polymerase. These three short ds DNAs were subsequently mixed together in a ratio of about 3∶1∶3 and amplified by PCR using primers derived from 5’end of the S1 and 3’end of the S6. The artifical gene fragment was thus produced largely. Addition of restriction sites to the primers allowed easy cloning of the artifical gene fragments.更多还原