Unit

UNIT 10.24 cDNA Amplification Using One-Sided (Anchored) PCR

  1. Robert L. Dorit1,
  2. Osamu Ohara2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.im1024s08

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Dorit, R. L. and Ohara, O. 2001. cDNA Amplification Using One-Sided (Anchored) PCR. Current Protocols in Immunology. 8:VII:10.24:10.24.1–10.24.10.

Author Information

  1. 1

    Yale University, New Haven, Connecticut

  2. 2

    Shionogi Research Laboratories, Osaka, Japan

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: DEC 1993

Abstract

This unit presents a modification of PCR, called anchored PCR, that allows amplification of full-length mRNA when only a small amount of sequence information lying within the mRNA is available. Both the original and reamplifications use an oligo(dT) primer complementary either to the poly(A) tail of the mature mRNA [when amplifying downstream (3') to the known sequence] or to an enzymatically synthesized homopolymer tail added to the cDNA following first strand synthesis [when amplifying upstream (5') to the known sequence]. The two rounds of PCR amplification result in a single product that can be sequenced directly or cloned into an appropriate vector for further analysis.