PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time)—eliminate genomic DNA contamination
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.
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.
Overview
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Genomic DNA contamination elimination in only 2 minutes
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Fast cDNA synthesis—reverse transcription in just 15 minutes
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Suitable for use with routine and challenging RNA templates alike
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Full-length cDNA synthesis up to 12 kb
More Information
Applications
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Elimination of genomic DNA contamination
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cDNA synthesis for real-time RT-PCR (qPCR)
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Two-step real-time RT-PCR
Choose the right PrimeScript kit for real-time RT-PCR:
| If you want to.... | Use this.... |
| Make cDNA using a master mix that includes RTase and primers | PrimeScript RT Master Mix (Perfect Real Time) (Cat. # RR036A and RR036B) |
| Make cDNA using a kit that provides RTase and primers as separate components | PrimeScript RT Reagent Kit (Perfect Real Time) (Cat. # RR037A and RR037B) |
| Avoid genomic DNA contamination during cDNA synthesis and RT-qPCR | PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time) (Cat. # RR047A and RR047B) |
| Perform 1-step real-time RT-PCR with TB Green | One-Step TB Green PrimeScript RT-PCR Kit II (Perfect Real Time) (Cat. # RR086A and RR086B) |
| Prepare cDNA for RT-qPCR directly from fewer than 1,000 cells | CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 (Cat. # 3734) |
Product citations
Fukui, K. et al. Selective mimics of strigolactone actions and their potential use for controlling damage caused by root parasitic weeds. Mol. Plant 6, 88–99 (2013).
Nakajima, A. et al. Lon protease of Azorhizobium caulinodans ORS571 is required for suppression of reb gene expression. Appl. Environ. Microbiol. 78, 6251–61 (2012).
Nemoto, T., Mano, A. & Shibasaki, T. Increased expression of miR-325-3p by urocortin 2 and its involvement in stress-induced suppression of LH secretion in rat pituitary. Am. J. Physiol. - Endocrinol. Metab. 302, (2012).
Qin, P. et al. ABCG15 encodes an ABC transporter protein, and is essential for post-meiotic anther and pollen exine development in rice. Plant Cell Physiol. 54, 138–154 (2013).
Sasaki, N. et al. The splice variant Ntr encoded by the tobacco resistance gene N has a role for negative regulation of antiviral defense responses. doi:10.1016/j.pmpp.2013.08.002
Sato, Y. et al. Anks4b, a novel target of HNF4α protein, interacts with GRP78 protein and regulates endoplasmic reticulum stress-induced apoptosis in pancreatic β-cells. J. Biol. Chem. 287, 23236–45 (2012).
Wang, Q. et al. Characterization of CYP76M5-8 indicates metabolic plasticity within a plant biosynthetic gene cluster. J. Biol. Chem. 287, 6159–68 (2012).
Xiang, J.-J., Zhang, G.-H., Qian, Q. & Xue, H.-W. Semi-rolled leaf1 encodes a putative glycosylphosphatidylinositol-anchored protein and modulates rice leaf rolling by regulating the formation of bulliform cells. Plant Physiol. 159, 1488–500 (2012).
Yang, Y. et al. Expression of the laccase gene from a white rot fungus in Pichia pastoris can enhance the resistance of this yeast to H2O2-mediated oxidative stress by stimulating the glutathione-based antioxidative system. Appl. Environ. Microbiol. 78, 5845–54 (2012).
Zhou, M.-L. et al. Genome-wide identification of genes involved in raffinose metabolism in Maize. Glycobiology 22, 1775–1785 (2012).
Zhou, Z. et al. Follicular development and expression of nuclear respiratory factor-1 and peroxisome proliferator-activated receptor coactivator-1 alpha in ovaries of fetal and neonatal doelings. J. Anim. Sci. 90, 3752–3761 (2012).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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