LIGATION-ANCHORED PCR - A SIMPLE AMPLIFICATION TECHNIQUE WITH SINGLE-SIDED SPECIFICITY
- Author(s)
- TROUTT, AB; MCHEYZERWILLIAMS, MG; Pulendran, B; Nossal, GJV;
- Details
- Publication Year 1992-10-15,Volume 89,Issue #20,Page 9823-9825
- Journal Title
- PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
- Publication Type
- Journal Article
- Abstract
- A simple, efficient, and sensitive technique has been developed for amplification of cDNAs encoding molecules with 5' regions of unknown sequence. In this ligation-anchored PCR, T4 RNA ligase is used to covalently link an "anchor" oligonucleotide to first-strand cDNAs. These anchored cDNAs are then amplified by using one PCR primer specific for the anchor and another specific for a sequence within the molecule of interest. The anchor oligonucleotide has been especially designed to facilitate subsequent analysis and cloning of the resultant PCR products. This three-stage procedure does not require purification of product between steps and avoids many of the technical difficulties associated with established anchored PCR protocols. The efficacy of ligation-anchored PCR was demonstrated by amplification of a specific IgG1 cDNA; total RNA equivalent to as few as 100 cells yielded the expected PCR product.
- Publisher
- NATL ACAD PRESS
- Keywords
- POLYMERASE CHAIN-REACTION; CELLS; GENES
- Publisher's Version
- https://doi.org/10.1073/pnas.89.20.9823
- Terms of Use/Rights Notice
- Refer to copyright notice on published article.
Creation Date: 1992-10-15 12:00:00