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Efficient and Reliable PCR Setup Using Eppendorf MasterMix

Christian Rohrer and Lars-Erik Peter
Christian Rohrer, Eppendorf AG, Germany
Lars-Erik Peters, Eppendorf 5Prime, USA

Introduction

The reliability of a PCR* experiment is dependent on its sensitivity, specificity and reproducibility. An essential prerequisite for a successful PCR is efficient sample preparation, as well as thermal cyclers with excellent temperature precision and block homogeneity.

Preparing a PCR reaction using different reaction components is afforded a particularly crucial role since the precision of the reagent concentrations from tube to tube, as well as from one experiment to the next, must be guaranteed. When a PCR preparation is being carried out, a series of comprehensive pipetting steps are necessary. It is advisable to minimize the number of pipetting steps as every additional step is accompanied by the risk of reduced precision and increased contamination.

Only constant concentration of the reaction components from one preparation to the next can guarantee the best possible reproducibility of the individual reactions. Ready-to-use MasterMix eliminates the need for time-consuming pipetting steps, since only primer and template need to be added to 2.5 x MasterMix. This not only minimizes the risk of errors, but it also optimizes precision, as well as saves a great deal of time.

In addition to reagents such as Taq polymerase, nucleotides and a buffer system, MasterMix contains special stabilizers, which ensure high performance in the PCR and excellent stability of the reaction components.

Objective of the Test

Eppendorf MasterMix was tested under various conditions. The o
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