Cells to cDNA: CellAmp whole transcriptome amplification kit for RT-PCR
The CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 enables you to go from cells to cDNA that is ready for analysis by real-time PCR. When using conventional techniques for working with trace amounts of nucleic acids, loss of product during purification steps is a frequent problem. However, this cells-to-cDNA kit ensures good product yield because loss-inducing purification steps for RNA and cDNA are not required.
The CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 enables you to go from cells to cDNA that is ready for analysis by real-time PCR. When using conventional techniques for working with trace amounts of nucleic acids, loss of product during purification steps is a frequent problem. However, this cells-to-cDNA kit ensures good product yield because loss-inducing purification steps for RNA and cDNA are not required.
The procedure begins with cell lysis, followed by a reverse transcription reaction with the dT adapter primer (RT dT Primer 2) to synthesize cDNA from mRNA. Next, terminal deoxynucleotidyl transferase (TdT) is used to add a dA tail to the synthesized cDNA. The resulting 3' dA tail containing cDNA product is used as direct template for PCR, resulting in fewer required amplification cycles.
In addition to moving from cells to cDNA, this product can be used to amplify cDNA from trace amounts of RNA.
The CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 uses an additional exonuclease I treatment step, which helps suppress nonspecific primer-derived amplification and increases yields of low-expressed genes.
Overview
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Allow direct amplification of cDNA from a small number of cultured cells (<1,000) using real-time RT-PCR
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Allows real-time PCR (qPCR) analysis of multiple genes from small samples
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Expression analysis for low-expressed gene
More Information
Applications
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Cells to cDNA with minimal purification
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Real-time RT-PCR (RT-qPCR)
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cDNA amplification from trace amounts of RNA
| Choosing a PrimeScript kit for real-time RT-PCR | |
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If you want to… |
Use this… |
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Make cDNA using a master mix that includes RTase and primers |
PrimeScript RT Master Mix (Perfect Real Time) (Cat. # RR036A and RR036B) |
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Make cDNA using a kit that provides RTase and primers as separate components |
PrimeScript RT Reagent Kit (Perfect Real Time) (Cat. # RR037A and RR037B) |
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Avoid genomic DNA contamination during cDNA synthesis and RT-qPCR |
PrimeScript RT Reagent Kit with gDNA Eraser (Perfect Real Time) (Cat. # RR047A and RR047B) |
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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) |
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Prepare cDNA for RT-qPCR directly from fewer than 1,000 cells |
CellAmp Whole Transcriptome Amplification Kit (Real Time), Ver. 2 (Cat. # 3734) |
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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