侧翼序列克隆方法评价

Estimation of Techniques about Cloning the Flanking DNA Sequence

克隆已知序列的侧翼DNA区域是基因操作实验中经常面临的任务之一。自PCR技术发明以来,以其为基础克隆侧翼序列的方法不断开发,如反向PCR(inversePCR)、捕捉PCR(capturePCR)、VectorettePCR、抑制PCR(suppressionPCR)、T接头PCR(T-linkerPCR)、多功能接头PCR(versatilecassettePCR)、AluPCR、热不对称交错PCR(thermalasymmetricinterlacedPCR)和DNA步行—复性控制引物(DNAwalking-an-nealingcontrolprimerTM)等。本文对上述方法的原理进行了简要的概括和总结,据此将其归为三大策略,即连接成环PCR、外源接头介导PCR和半随机引物PCR。对不同的策略和同一策略中不同的方法的优缺点亦进行了比较,进而讨论和展望了它们的主要应用领域和潜力,以期对实际操作起到借鉴作用。

Cloning DNA fragments flanking the known sequences is frequently involved in many DNA manipulation experiments. Since the invention of PCR technique, adaptation techniques to isolate flanking region continue to be developed and become the most important ways, such as inverse PCR, capture PCR, vectorette PCR, suppression PCR, T-linker PCR, versatile cassette PCR, Alu PCR, thermal asymmetric interlaced PCR and DNA Walking-Annealing Control Primer^TM. In this review, the principles of these techniques were briefly outlined and compared. Based on this, these techniques were grouped into three different strategies, for example, orbicular-ligation PCR, adapter-mediated PCR and semi-random primer PCR. After analyzing the advantages and disadvantages of these different strategies or techniques in the same strategies, we discussed the application field and development prospect of corresponded techniques.