Open AccessHighly Sensitive Method to Detect mRNAs in Individual Cells by Direct RT-PCR Using Tth DNA Polymerase
Author(s) -
Gilles Chiocchia,
Kendall A. Smith
Publication year - 1997
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/97222st04
Subject(s) - microbiology and biotechnology , biology , real time polymerase chain reaction , polymerase chain reaction , reverse transcription polymerase chain reaction , dna , dna polymerase , reverse transcriptase , messenger rna , gene , genetics
A new method for detecting the expression of low-abundance mRNA molecules has been developed that combines the sensitivity of PCR, the high efficiency and specificity of reverse transcription (RT) using Tth DNA polymerase at high temperature, and the enhancement of sensitivity and specificity of nested PCR. This method is highly sensitive, reproducible and allows the detection of mRNAs in individual cells by direct RTPCR.