UNIT 14.8 In situ Polymerase Chain Reaction and Hybridization to Detect Low-Abundance Nucleic Acid Targets
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- Published Print: APR 1996
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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.