Cells-to-CT™ 1-Step TaqMan® Kit

Catalog number: A25605

Ambion™  Related applications: Real Time PCR-Based Gene Expression Profiling

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Description

Like other members of the Cells-to-CT™ product family, the Cells-to-CT™ 1-Step TaqMan® Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification.

This new addition to the family includes a TaqMan® 1-Step RT-PCR master mix. The single-tube format of the RT-PCR step streamlines the “sample to answer” workflow more than ever. Combining the simple Cells-to- CT™ lysis technology with the fast cycling capability of the 1-step RT-PCR master mix allows quicker time to results with limited sample handling.

The Cells-to-CT™ 1-Step TaqMan® Kit is:
• Complete—optimized workflow includes cell lysis reagents with gDNA removal and TaqMan® 1-Step RT-PCR Master Mix
• Faster than ever—7-minute sample prep, including DNase treatment, combined with a 28-minute real-time RT-PCR step allows for ~35 minute sample-to-answer workflow
• Simple—10 steps total (sample prep and real-time RT-PCR combined)
• Robust—perform gene expression analysis on 10–100,000 cells per sample; results equivalent to those from purified RNA
• Convenient—contains sufficient reagents to perform 5 real-time RT-PCRs for each cell sample you prepare; this allows you to study multiple targets or perform replicate RT-PCRs

Note: TaqMan® primer/probe sets are sold separately.

Simple 7-minute Sample Preparation, Then Ready for Real-time RT-PCR
Whether you are using plates or tubes, the Cells-to-CT™ 1-Step TaqMan® Kit uses a simple 7-minute sample preparation procedure. The lysis technology is designed for 10–100,000 cultured cells/sample. Cells are washed in PBS and lysed in solution for 5 minutes at room temperature with a simultaneous DNase treatment. Lysis is terminated at room temperature by a 2-minute incubation with Stop Solution. The lysate is now ready to analyze on your real-time PCR instrument. Simply add it to a real-time RT-PCR master mix containing the TaqMan® primer/probe sets of interest. Alternatively, the lysate can be stored at –20°C. Because samples can be processed directly in culture wells (96 or 384 wells), sample handling and the potential for sample loss or transfer error are minimized, facilitating rapid, high-throughput processing. Unlike old-fashioned, multi-step RNA isolation protocols, no heating, washing, or centrifugation steps are required. The kit greatly simplifies a laborious 30–60 minute process and reduces it to 7 minutes.

A Complete, Validated Solution
All components of the Cells-to-CT™ 1-Step TaqMan® Kit have been optimized for consistent and reliable performance. This removes the guesswork involved in assembling separate sample preparation and real-time 1-step RT-PCR master mixes. For added quality assurance, the Cells-to-CT™ 1-Step TaqMan® Kit has been validated with TaqMan® Gene Expression assays and shows performance equivalent to that obtained with purified RNA.

Accessory Product
The TaqMan® Cells-to-CT™ Control Kit was designed for use with TaqMan® Cells-to-CT™ products. This kit contains an endogenous control (ACTB) to normalize for sample loading variability and an artifical XenoRNA™ control and corresponding TaqMan® Gene Expression assay to monitor the effects of PCR inhibition.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

PCR Method: 1-Step qRT-PCR
Detection Method: Primer-Probe Detection
Reaction Speed: Standard, Fast
For Use With (Equipment): 7900HT System, 7500 System, 7900HT Fast System, StepOnePlus™ System, 7500 Fast Dx System, ViiA™ 7 System, 7500 Fast System, QuantStudio™, StepOne™ System, Real-Time PCR Systems or Thermal Cyclers
Sample Type (General): RNA, Cells
Product Size: 20 preps

Contents & storage

• Lysis Solution, 2.2 mL, store at 2–8°C
• Dnase I, 22 µL, store at -5 to -30°C
• Stop Solution, 200 µL, store at -5 to -30°C
• TaqMan® 1-Step qRT-PCR Mix, 250 µL, store at -5 to -30°C

Documents

Manuals & protocols

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