UNIT 10.24 cDNA Amplification Using One-Sided (Anchored) PCR
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- Published Print: DEC 1993
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.