Ambion® Cells-to-CT™ Kits
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Read What Our Customers Have to Say
"The TaqMan Gene Expression Cells-to-Ct Kit has performed consistently. The performance is increased as compared to RNA purification, RT steps and then qRT-PCR. There is less handling, less chance at mistakes. Because of this time savings and increase in sensitivity, I've incorporated this protocol into the Human Embryonic Stem Cell Training Course at the Stem Cell Research Center at Rutgers University (http://www.stemcellcourse.org)."
Dr. Rick Cohen, Director, hESC Core Facility, Stem Cell Research Center,
Rutgers University, Piscataway, NJ
Overview
Simple, Complete Workflows for Gene Expression Analysis without RNA Purification
Cells-to-CT™ Kits allow you to directly get real-time RT-PCR results from cells, without having to purify RNA. These kits provide unsurpassed performance in gene expression and microRNA analysis, including detection of siRNA-induced mRNA knockdown. Cells are simply lysed at room temperature in ≤10 minutes; an optional DNase digestion can be performed concurrently during lysis.
Cells to CT™ vs Kits from Alternative Vendors
Additionally, the performance of the TaqMan Gene Expression Cells-to-CT Kit was compared to lysate kits from alternative vendors and to purified RNA. Inputs of 100 - 100,000 cells/lysis reaction were examined. The sensitivity of the TaqMan Cells-to-CT Kit protocol was equivalent to that obtained with purified RNA, and it surpassed those from alternative vendors (Figure 1).
 | Figure 1: The Sensitivity of the TaqMan® Gene Expression: HeLa cells (100–100,000) were prepared using either traditional RNA purification, the TaqMan® Gene Expression Cells-to-CT™ Kit protocol, or other lysate methods from alternative vendors Q and S. Reverse transcription reactions comprised the maximum recommended sample input for each kit, and real-time PCR was performed in triplicate using a PPIA TaqMan G 15 ene Expression Assay. |
Equal or Better Performance & Sensitivity Compared to Purified RNA
 | Figure 2: Detection of limited target sequences. Increasing amounts of NIH3T3 cells (mouse) were added to a constant 10,000 HeLa cells (human). These cells were processed in triplicate by the TaqMan® Gene Expression Cells-to-CT™ Kit or purified reagents provided using a RNA spin column method. All samples were reverse transcribed by the RT for mousespecific (mM B2M) and human-specific (Hs. TKT) genes on all samples in triplicate reactions on a 7900HT Fast Real-Time PCR System. |
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Featured Articles
- Tips & Tricks for Using Ambion® Cells to Ct
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- Human Embryonic Stem Cell Gene Expression and Cell Fate
- Mature miRnA Analysis Directly from Cell Lysates
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- Real-Time RT-PCR Directly from Cell Lysates
- Don’t Bother Quantitating RNA for Gene Expression Analysis
- Automate Cells-to-CT™ Kits for High Throughput Expression Analysis
- Variation of microRNA Profiles in Normal vs. Cancerous Hepatocytes
- Go from Cells in Culture to Real-Time PCR Data Even Faster
- Making the Most of Limited Samples: Skip RNA Isolation and PreAmplify Cell Lysates Directly
- Ideal Control for PCR Inhibition and Cell Input
- siRNA Screening: Validate Thousands of Targets in a Single Week
- Using SYBR® Green for Real-Time RT-PCR? Try SYBR Green Cells-to-CT™ Kits
- Real-Time RT-PCR Directly from Cell Lysates: A Complete Workflow without RNA Purification
