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. 1992 Oct 15;89(20):9823–9825. doi: 10.1073/pnas.89.20.9823

Ligation-anchored PCR: a simple amplification technique with single-sided specificity.

A B Troutt 1, M G McHeyzer-Williams 1, B Pulendran 1, G J Nossal 1
PMCID: PMC50225  PMID: 1409706

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.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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