Unit

UNIT 25B.5 AFLP-Based Transcript Profiling

  1. Pieter Vos,
  2. Patrick Stanssens

Published Online: 1 FEB 2002

DOI: 10.1002/0471142727.mb25b05s57

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Vos, P. and Stanssens, P. 2002. AFLP-Based Transcript Profiling. Current Protocols in Molecular Biology. 57:B:25B.5:25B.5.1–25B.5.16.

Author Information

  1. Keygene N.V., The Netherlands

Publication History

  1. Published Online: 1 FEB 2002
  2. Published Print: JAN 2002

Abstract

This unit presents an alternative to differential display that allows the quantification of transcripts, based on AFLP-fingerprinting of double-stranded cDNA. The protocol described includes the following steps: the isolation of poly(A)+ RNA from total RNA, the synthesis of double-stranded cDNA, the preparation of template fragments by digestion of the cDNA library with a combination of two restriction enzymes and the ligation of adaptors to the fragment ends, the selective amplification of specific subsets of fragments, and the electrophoretic analysis of these amplification products on standard denaturing polyacrylamide gels. The transcript profiles obtained by this technique are a reliable and efficient tool to identify differentially expressed mRNAs.This unit presents an alternative to differential display that allows the quantification of transcripts, based on AFLP-finger his unit presents an alternative to differential display that allows the quantification of transcripts, based on AFLP-finger