RAPID AND EFFICIENT COSMID CLONING
RAPID AND EFFICIENT COSMID CLONING
复制标题
DOI：
10.1093/nar/9.13.2989
复制
发表时间：
1981-01-01
影响因子：
14.9
通讯作者：
BURKE, JF
中科院分区：
文献类型：
--
作者：
ISHHOROWICZ, D;BURKE, JF
We present a procedure for cosmid cloning that allows rapid and efficient cloning of individual DNA fragments of between 32kb and 45kb. By appropriate treatment of the cloning vector, pJB8, we make left-hand and right-hand vector ends that are incapable of self-ligation but which accept dephosphorylated insert DNA fragments. The inserted fragments are generated by partial digestion withMboI or Sau3A and are dephosphorylated to prevent ligation and insertion of non-contiguous fragments. The method eliminates the need to size the insert DNA fragments and prevents formation of clones containing short or multiple inserts. 1μg of targetDrosophilaDNA gives about 5×105clones, with an average insert size of 38kb. We also describe a rapid and efficient method for preparing plasmid and cosmid DNA.
