Genomic DNA contamination can be a significant problem during real-time RT-PCR because co-amplification of genomic DNA and cDNA can lead to erroneous results. With the PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time), genomic DNA contamination is eliminated. This reverse transcription kit enables fast cDNA synthesis for real-time RT-PCR (qPCR) and includes a rapid step that eliminates genomic DNA (gDNA) contamination without loss of input RNA.

The PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time) includes PrimeScript reverse transcriptase, a modified RNase H-minus recombinant MMLV (Moloney Murine Leukemia Virus) enzyme which allows fast cDNA synthesis due to its exceptionally strong strand-displacement activity. PrimeScript RTase is robust, versatile, and can be used to prepare cDNA from almost any RNA template, including GC-rich templates and RNAs with significant secondary structure. Because of the excellent extension capacity of PrimeScript, cDNA synthesis reactions can be performed at lower temperatures (i.e., 42°C), decreasing the risk of RNA degradation.

In the first step, genomic DNA contamination is eliminated by treating the samples with gDNA Eraser (an optimized, potent DNA-degradation enzyme) for 2 minutes at 42°C. The subsequent reverse transcription reaction includes a component that completely inhibits DNA degradation activity, and reverse transcription is complete in just 15 minutes. The final result is high-quality cDNA with no trace of genomic DNA contamination. This kit is designed for two-step RT-PCR and should be used with one of Takara Bio"s qPCR premixes such as TB Green Premix Ex Taq II (Tli RNase H Plus) (Cat. # RR820A), TB Green Premix Ex Taq (Tli RNase H Plus) (Cat. # RR420A), or Premix Ex Taq (Probe qPCR) (Cat. # RR390A).

Over 3,700 peer-reviewed articles have been published in which PrimeScript reverse transcriptase is applied towards the study of gene expression, gene discovery, transcriptome analysis, and miRNA expression regulation.