PrimeScript RT Reagent Kit (Perfect Real Time)—generate cDNA for real-time PCR
When synthesizing cDNA for real-time PCR, the PrimeScript RT Reagent Kit (Perfect Real Time) allows maximal flexibility during reaction assembly. This reverse transcription reagent kit facilitates cDNA synthesis prior to analysis by real-time PCR. Designed for two-step real-time RT-PCR (also called qRT-PCR or RT-qPCR), the PrimeScript RT Reagent Kit (Perfect Real Time) contains all of the components needed for reverse transcription, including PrimeScript RTase, RNase inhibitor, random 6-mers, oligo dT primer, dNTPs, and reaction buffer. Because the kit's primers and RTase are provided as separate components, there is greater flexibility in reaction assembly.
When synthesizing cDNA for real-time PCR, the PrimeScript RT Reagent Kit (Perfect Real Time) allows maximal flexibility during reaction assembly. This reverse transcription reagent kit facilitates cDNA synthesis prior to analysis by real-time PCR. Designed for two-step real-time RT-PCR (also called qRT-PCR or RT-qPCR), the PrimeScript RT Reagent Kit (Perfect Real Time) contains all of the components needed for reverse transcription, including PrimeScript RTase, RNase inhibitor, random 6-mers, oligo dT primer, dNTPs, and reaction buffer. Because the kit's primers and RTase are provided as separate components, there is greater flexibility in reaction assembly.
The PrimeScript RT Reagent Kit (Perfect Real Time) includes PrimeScript reverse transcriptase for strong strand-displacement activity and efficient synthesis of cDNA for real-time PCR. PrimeScript RTase is a modified, recombinant MMLV (Moloney murine leukemia virus) reverse transcriptase and is verified to be RNase H-minus. PrimeScript RTase is also robust, versatile, and can be used for reverse transcription of virtually any RNA template, including templates that have a high degree of secondary structure or are GC rich. Because of the excellent extension capability of PrimeScript, cDNA synthesis reactions can be performed at 42°C, minimizing the risk of RNA degradation that often occurs at higher temperatures.
If a master mix is preferred, consider using PrimeScript RT Master Mix (Perfect Real Time). Use this kit in combination with one of Takara Bio's qPCR premix products, 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), for intercalating green dye detection or 5'-nuclease probe-mediated assays.
Overview
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Allows fast, efficient synthesis of cDNA for real-time PCR. This kit is designed for two-step real-time RT-PCR.
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Includes random 6-mers and oligo dT primer for use as reverse transcription primers. The reaction can be performed using one or a mixture of these two primer types; alternately, gene-specific primers may be used.
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Includes EASY Dilution (for Real Time PCR) when generating reference standards for a qPCR standard curve. This reagent facilitates higher accuracy at lower standard concentrations.
More Information
Applications
- cDNA synthesis for real-time 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) |
Note
A standard curve must be generated for quantitation by real-time PCR. Dilution of total RNA or cDNA is necessary in order to include low concentration samples, which are important for a viable standard curve with an appropriately wide range. However, dilution with water or TE can narrow the range of the curve because nucleic acids are unstable at low concentrations in the presence of dH2O or TE. Use EASY Dilution (for Real Time PCR) provided in this kit for generating accurate reference standards.
Product citations
Zha, X. et al. Lactate Dehydrogenase B Is Critical for Hyperactive mTOR-Mediated Tumorigenesis. Cancer Res. 71, (2011).
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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