Rapid and efficient cosmid cloning

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ID: 291165
1981
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Abstract
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 dephosporylated insert DNA fragments. The inserted fragments are generated by partial digestion with MboI 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. 1 microgram of target Drosophila DNA gives about 5 x 10(5) clones, with an average insert size of 38kb. We also describe a rapid and efficient method for preparing plasmid and cosmid DNA.
Reference Key
openalex_W2049635421 Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors David Ish‐Horowicz, J. F. Burke
Journal Nucleic Acids Research
Year 1981
DOI
10.1093/nar/9.13.2989
URL
https://doi.org/10.1093/nar/9.13.2989
Keywords Keywords not found

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