Open AccessSuccessful gene expression analysis by multiplex, real-time, one-step RT-PCR, irrespective of the targets amplified
Author(s) -
Holger Engel,
Corinna Kueppers,
Margaretha Koenig,
Dirk Loeffert
Publication year - 2007
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 - 10.2144/000112560
Subject(s) - multiplex , gene expression , biology , real time polymerase chain reaction , gene , microbiology and biotechnology , multiplex polymerase chain reaction , genetics , polymerase chain reaction , computational biology
Multiplex, real-time, one-step RT-PCR enables simultaneous quantification of several transcripts in one reaction vessel. This saves time and conserves precious sample material when analyzing the expression of reference and target genes in a particular sample. However, intensive PCR optimization is often required ( 1 ). We show that success in multiplex, real-time, one-step RT-PCR can be achieved at the first attempt with the QuantiTect® Multiplex RT-PCR Kit, regardless of the combination of genes analyzed. In each of the multiplex assays carried out, all targets were amplified with high efficiency, which is important for precise relative quantification of gene expression levels by the ΔΔC T method.