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This application describes the use of a new commercial PCR enhancer for improving PCR efficiency: the TaqMaster. With this enhancer, an integral component of the MasterTaq Kit by Eppendorf, it was possible to amplify human hsp60 (heatshock protein) and hsp27 amplicons, an application in which five other polymerases, including hot start and proofreading enzymes, failed. The application of the PCR enhancer TaqMaster is highly recommended for solving problems with difficult targets, for detecting low copy RNAs or for increasing product yield.
Introduction
Reverse Transcription Polymerase Chain Reaction (RT-PCR) is a common
method for the detection of gene expression. It can be used qualitatively
to show principle expression of a specific target gene in a cell type
or it may even serve to quantify the expression level of the gene of interest.
In differential gene expression analysis, a search is undertaken for target
genes that are regulated by certain conditions, eg, application of drugs.
There are several methods for identifying these regulated genes; differential
display RT-PCR,1 subtractive hybridization2 or complex hybridization onto
cDNA arrays3 are some of the most common. With these techniques, a set
of candidate sequences is obtained with or without a known identity. All
these potentially regulated sequences need further confirmation that they
really have been expressed different
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