Unit

UNIT 14.8 In situ Polymerase Chain Reaction and Hybridization to Detect Low-Abundance Nucleic Acid Targets

  1. Omar Bagasra1,
  2. Thikkavarapu Seshamma1,
  3. Roger Pomerantz1,
  4. John Hanson2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1408s34

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Bagasra, O., Seshamma, T., Pomerantz, R. and Hanson, J. 2001. In situ Polymerase Chain Reaction and Hybridization to Detect Low-Abundance Nucleic Acid Targets. Current Protocols in Molecular Biology. 34:14.8:14.8.1–14.8.24.

Author Information

  1. 1

    Thomas Jefferson University, Philadelphia, Pennsylvania

  2. 2

    MJ Research, Watertown, Massachusetts

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 1996

This is not the most recent version of the article. View current version (1 APR 2008)

Abstract

This unit presents a novel approach for detecting low-abundance nucleic acid targets in nuclear and cytoplasmic regions by amplification of specific target sequences using an in situ polymerase chain reaction (ISPCR). If the target sequence is RNA, ISPCR is preceded by in situ reverse transcription. Following ISPCR, in situ hybridization is performed. An Alternate Protocol describes a variant in situ method for simultaneously reverse transcribing and amplifying RNA transcripts using recombinant Thermos thermophilos (rTth) polymerase. In situ hybridization and detection of amplified targets is also described.