( ...)Enhancing PCR* Efficiency with MasterTaq

Bodo Eickhoff and Jrgen van der Bosch
Experimental Immunopharmacology, Research Center Borstel, Parkallee 22, D-23845 Borstel, Germany.
Tel.: +49 4537 188458 Fax: +49 4537 188404
email: jvdbosch@fz-borstel.de

Abstract

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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