Linked Life Data

1367662

Source:http://linkedlifedata.com/resource/pubmed/id/1367662

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
7
pubmed:dateCreated
1992-1-2
pubmed:abstractText
We have devised a simple, universal cloning strategy that permits the direct ligation of PCR amplified nucleic acid to a compatible vector preparation. The method does not require that special restriction sites or additional sequences be appended onto the amplification primers, nor the use of restriction endonucleases, modifying enzymes, or any purification procedures. This approach takes advantage of the single 3' deoxyadenylate extension that Thermus aquaticus, Thermus flavus, and Thermococcus litoralis DNA polymerases add to the termini of amplified nucleic acid. A new type of plasmid was constructed that allows the preparation of ends containing a single complementary 3' deoxythymidylate extension. Cloning PCR products by this method is approximately 50 times more efficient than blunt-ended ligation reactions. This direct PCR ligation technique has been engineered in-phase with a truncated lacZ gene to facilitate the discrimination of recombinants from non-recombinants. We have applied this method to directly clone amplified RNA and DNA from as little as 1 microliter of a PCR reaction, without prior modification or purification steps.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
B
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0733-222X
pubmed:author
pubmed:issnType
Print
pubmed:volume
9
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
657-63
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed:year
1991
pubmed:articleTitle
A universal method for the direct cloning of PCR amplified nucleic acid.
pubmed:affiliation
Department of Chemistry, University of Wisconsin, Madison 53706.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't