Chain reaction cloning: a one-step method for directional ligation of multiple DNA fragments

被引：26

作者：

Pachuk, CJ

Samuel, M

Zurawski, JA

Snyder, L

Phillips, P

Satishchandran, C

机构：

[1] Wyeth Ayerst Res, Malvern, PA 19355 USA

[2] Univ Penn, Sch Vet Med, Dept Pathobiol, Philadelphia, PA 19104 USA

[3] Centocor Inc, Malvern, PA 19355 USA

来源：

GENE | 2000年 / 243卷 / 1-2期

关键词：

bridge oligonucleotides; multifragment ligation; sequence-specific ligation; site-specific ligation; thermostable ligation;

D O I：

10.1016/S0378-1119(99)00508-9

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

A novel DNA assembly method, chain reaction cloning (CRC), is described. CRC enables the ordered assembly of multiple DNA fragments in a single step. The power of the technique was demonstrated by the directed in vitro assembly of a plasmid comprised of six DNA fragments from a pool of 12 available fragments. The odds of obtaining the correct plasmid clone in a single step, using conventional techniques, is less than 1 in 191 000 000. Using CRC, the desired plasmid was recovered at a frequency of one in two. Ligation is no longer the rate limiting step in cloning, and limitless possibilities exist for the reconstruction of complex genomes. (C) 2000 Elsevier Science B.V. All rights reserved.

引用

页码：19 / 25

页数：7