rdf:type |
|
lifeskim:mentions |
|
pubmed:issue |
2
|
pubmed:dateCreated |
1995-8-28
|
pubmed:abstractText |
A reliable selective PCR procedure that combines the use of additionally mutated primers with the specificity-enhancing properties of a commercial preparation (Perfect Match, Stratagene) is described. The human immunodeficiency virus type 1 pol gene point mutations known to confer in vitro resistance to azidothymidine were examined as a model for optimization of the assay. The usual strategy of deliberately introducing an additional mismatch 1 residue from the 3' end in the wild-type and mutant primers did not allow reproducible discrimination between wild-type and mutant target sequences. Addition of minimal amounts of Perfect Match to the same PCR mixtures resulted in a significantly enlarged range of selective annealing temperatures, providing a valuable and cost-effective means for reliable detection of known mutations by selective PCR.
|
pubmed:language |
eng
|
pubmed:journal |
|
pubmed:citationSubset |
IM
|
pubmed:chemical |
|
pubmed:status |
MEDLINE
|
pubmed:month |
Apr
|
pubmed:issn |
1073-6085
|
pubmed:author |
|
pubmed:issnType |
Print
|
pubmed:volume |
3
|
pubmed:owner |
NLM
|
pubmed:authorsComplete |
Y
|
pubmed:pagination |
166-9
|
pubmed:dateRevised |
2006-11-15
|
pubmed:meshHeading |
pubmed-meshheading:7620977-Base Sequence,
pubmed-meshheading:7620977-DNA,
pubmed-meshheading:7620977-DNA Primers,
pubmed-meshheading:7620977-DNA-Directed DNA Polymerase,
pubmed-meshheading:7620977-Drug Resistance, Microbial,
pubmed-meshheading:7620977-Genes, pol,
pubmed-meshheading:7620977-HIV-1,
pubmed-meshheading:7620977-Humans,
pubmed-meshheading:7620977-Molecular Sequence Data,
pubmed-meshheading:7620977-Point Mutation,
pubmed-meshheading:7620977-Polymerase Chain Reaction,
pubmed-meshheading:7620977-Reproducibility of Results,
pubmed-meshheading:7620977-Taq Polymerase,
pubmed-meshheading:7620977-Temperature,
pubmed-meshheading:7620977-Zidovudine
|
pubmed:year |
1995
|
pubmed:articleTitle |
Increased reliability of selective PCR by using additionally mutated primers and a commercial Taq DNA polymerase enhancer.
|
pubmed:affiliation |
Dipartimento di Biologia Molecolare, Università di Siena, Italy.
|
pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
|