Cells-to-CT™ 1-Step Power SYBR® Green Kit

Catalog number: A25599

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 Power SYBR® Green Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification.

This new addition to the family includes a Power SYBR® Green 1-Step RT-PCR master mix. The 1-step format of the RT-PCR step streamlines the Cells-to- CT™ "sample to answer" workflow more than ever without compromising sensitivity and specificity and at an extraordinary value.

The Cells-to-CT 1-Step Power SYBR® Green Kit is:
• Complete—optimized workflow includes cell lysis reagents with gDNA removal and Power SYBR® Green 1-Step RT-PCR Master Mix
• Faster—7-minute sample prep, including DNase treatment, versus 30-60 minutes for traditional RNA purification
• Simple—12 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
• Affordable—contains sufficient reagents to perform 5 real-time RT-PCRs for each cell sample prepared; study multiple targets per sample at an affordable price

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 Power SYBR® Green 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 primer sets for your genes 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.

Completely Validated, Completely Affordable
All components of the Cells-to-CT™ 1-Step Power SYBR® Green 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 Power SYBR® Green Kit has been validated with a set of primers for common gene targets and shows performance equivalent to that obtained with purified RNA.

When you add it all up, the Cells-to-CT™ 1-Step Power SYBR® Green Kit is the best value for your gene expression experiments. With enough reagents to perform 5 real-time RT-PCRs per sample, the savings are substantial compared to buying master mix and an RNA purification kit separately to do the same work.

Accessory Product
The SYBR® Green Cells-to-CT™ Control Kit was designed for use with SYBR® Green Cells-to-CT™ products. This kit contains an endogenous control primer set (ACTB) to normalize for sample loading variability and an artifical XenoRNA™ control and corresponding primer set 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: SYBR
Reaction Speed: Standard
For Use With (Equipment): 7900HT System, StepOne™ System, Real-Time PCR Systems or Thermal Cyclers, QuantStudio™, StepOnePlus™ System, 7500 Fast Dx System, 7500 Fast System, 7900HT Fast System, 7500 System, ViiA™ 7 System
Sample Type (General): RNA, Cells
Product Size: 400 preps

Contents & storage

• Lysis Solution, 22 mL, store at 2–8°C
• Dnase I, 220 µL, store at -5 to -30°C
• Stop Solution, 2 x 1 mL, store at -5 to -30°C
• Power SYBR® Green 1-Step qPCR Mix, 20 mL, store at -5 to -30°C
• Power SYBR® Green 1-Step RT Mix, 320 µL, store at -5 to -30°C

Documents

Manuals & protocols

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