Unit

UNIT 21.19 The CLIP-CHIP Oligonucleotide Microarray: Dedicated Array for Analysis of All Protease, Nonproteolytic Homolog, and Inhibitor Gene Transcripts in Human and Mouse

  1. Reinhild Kappelhoff,
  2. Chris Overall

Published Online: 1 APR 2009

DOI: 10.1002/0471140864.ps2119s56

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Kappelhoff, R. and Overall, C. 2009. The CLIP-CHIP Oligonucleotide Microarray: Dedicated Array for Analysis of All Protease, Nonproteolytic Homolog, and Inhibitor Gene Transcripts in Human and Mouse. Current Protocols in Protein Science. 56:21.19:21.19.1–21.19.16.

Author Information

  1. University of British Columbia, Vancouver, British Columbia, Canada

Publication History

  1. Published Online: 1 APR 2009
  2. Published Print: APR 2009

Abstract

The CLIP-CHIP oligonucleotide microarray allows the analysis of mRNA transcript levels in a tissue sample for all proteases, nonproteolytic homologs, and protease inhibitors of the human and mouse genome. In the protocol presented in this unit, total RNA is extracted from a tissue, and the resulting mRNA is reverse transcribed into cDNA and dsDNA and then amplified in an in vitro transcription reaction. The amplified antisense RNA is labeled with a fluorescent dye and hybridized to the CLIP-CHIP, which contains unique oligonucleotides that are specifically designed for the protease, nonproteolytic homologs, protease inhibitors, and control samples. After hybridization, the fluorescence intensity of each spot is measured, thus identifying mRNA transcripts that are expressed and allowing basic quantification of expressed transcripts. Curr. Protoc. Protein Sci. 56:21.19.1-21.19.16. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • mouse and human oligonucleotide microarray;
  • linear amplification;
  • aRNA;
  • direct aRNA labeling