Self-sustained sequence replication (3SR): An alternative to PCR

James D. Mueller, Barbara Pütz, Heinz Höfler

Research output: Contribution to journalArticlepeer-review

25 Scopus citations

Abstract

The amplification of target nucleic acids before hybridization is one of the most powerful approaches for the detection of low copy number RNA and DNA. The best known amplification reaction is PCR which has many applications. However, certain drawbacks of the PCR reaction provide a role for alternative amplification methods. One of these methods is the self-sustained sequence replication (3SR) reaction, which is an isothermal method for RNA amplification depending on the action of three enzymes. 3SR has been used in several in vitro applications and has also been modified for in situ use (IS-3SR). We have studied IS-3SR with the measles virus as a model and have found that it can significantly amplify the amount of intracellular RNA. Such a level of amplification could raise the amount of single copy RNA to the level of detection by conventional in situ hybridization. Although careful controls to insure its specificity must be carried out, IS-3SR has several advantages, including ease of use, preserved cell morphology, and specificity for RNA amplification, which make it an attractive alternative to the in situ PCR method.

Original languageEnglish
Pages (from-to)431-437
Number of pages7
JournalHistochemistry and Cell Biology
Volume108
Issue number4-5
DOIs
StatePublished - 1997

Fingerprint

Dive into the research topics of 'Self-sustained sequence replication (3SR): An alternative to PCR'. Together they form a unique fingerprint.

Cite this