Efficient reamplification of differential display products by transient ligation and thermal asymmetric PCR

A new method for specific reamplification of DDRT-PCR products is presented. After transient ligation of the primary DDRT-PCR fragments into a T-vector, the cDNAs of interest were reamplified by hemi-nested PCR and thermally asymmetric cycles. In contrast to the originally described protocol, this m...

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Bibliographic Details
Published in:Nucleic acids research 1998-02, Vol.26 (4), p.1130-1131
Main Authors: Bonnet, S, Prevot, G, Bourgouin, C
Format: Article
Language:English
Subjects:
Animals
Anopheles - genetics
Anopheles - parasitology
Anopheles gambiae
Base Sequence
complementary DNA
DNA Ligases
DNA Primers - genetics
DNA, Complementary - genetics
Evaluation Studies as Topic
gene expression
Genetic Vectors
hematophagy
Life Sciences
messenger RNA
midgut
Nucleic Acid Amplification Techniques
Plasmodium falciparum - growth & development
Plasmodium falciparum - pathogenicity
polymerase chain reaction
Polymerase Chain Reaction - methods
Polymerase Chain Reaction - statistics & numerical data
Reproducibility of Results
Sensitivity and Specificity
Citations: Items that cite this one
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Description
Summary:A new method for specific reamplification of DDRT-PCR products is presented. After transient ligation of the primary DDRT-PCR fragments into a T-vector, the cDNAs of interest were reamplified by hemi-nested PCR and thermally asymmetric cycles. In contrast to the originally described protocol, this method of reamplification is specific, sensitive, reproducibly gives a high yield of DNA and allows direct sequencing of the reamplified product without purification or cloning.
ISSN:0305-1048
1362-4962
1362-4962
DOI:10.1093/nar/26.4.1130