Hot start polymerase chain reaction (PCR) is effortless using the FastStart™ PCR Master, a ready-to-use, double-concentrated hot start master mix. This master mix contains FastStart Taq DNA Polymerase, Roche high quality PCR-Grade nucleotides and all other reagents required for PCR and two-step reverse transcriptase (RT)-PCR on thermal cycler instruments. FastStart™ PCR Master accepts modified nucleotides such as Digoxigenin (DIG)-dUTP. Labeling with DIG-dUTP can be achieved by adding the modified nucleotide in a final concentration of 10 μM to the final volume. Combine this master mix with Roche′s Transcriptor First Strand cDNA Synthesis Kit to achieve excellent results in two-step RT-PCR.
Application
FastStart™ PCR Master has been used for the analysis of Nosema sp. DNA by polymerase chain reaction (PCR) method for validating the genomic DNA by PCR, for amplification of RNA by PCR method.
The FastStart™ PCR Master is a convenient alternative to the single reagents for nearly all hot start PCR applications. It can be applied for the amplification of genomic DNA and cDNA targets up to 2 kb long, or longer fragments on plasmid DNA targets, with high specificity, sensitivity, and yield. The enzyme is inactive at +15 to +25°C during PCR setup, and then activated at +95°C during initial denaturation. It can be used for: • Hot Start PCR up to 2 kb • Hot Start RT-PCR up to 2 kb • High-throughput PCR • Direct colony PCR