Power SYBR® Green Cells-to-CT™ Kit This product, and/or its packaging, incorporates one or more environmental/green features. Click on this icon to learn more.

Catalog number: 4402955

Ambion™  Related applications: Real Time PCR (qPCR) | Real Time PCR-Based Gene Expression Profiling

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Description

Ambion® Power SYBR® Green Cells-to-CT™ Kit is a fast, easy, and robust method to go directly from cultured cells to real-time PCR results. A breakthrough cell lysis and RNA stabilization technology eliminates the laborious and time-consuming RNA purification process completely. The kit includes sufficient reagents for 400 reactions. Power SYBR® Green Cells-to-CT™ Kit is Powered by AmpliTaq Gold® DNA Polymerase, LD, which provides the highest levels of specificity with standard real-time PCR.

• Includes optimized reagents to work efficiently, right out of the box
• Simple, effective Cells-to-CT™ technology enables sample preparation at room temperature in only 7 minutes, including DNase treatment
• Validated accuracy, exceptional reproducibility, and maximal sensitivity from 10 to 100,000 cells per sample; results equivalent to those from purified RNA

Extraordinary Value
The kit includes reagents for sample preparation integrated into a complete, optimized gene expression workflow including reverse transcription reagents and a high-performance PCR master mix with SYBR® Green.

Extraordinary Ease
The protocol begins with a simple 7-minute sample preparation procedure. First, cells are washed in PBS, and then lysed for 5 minutes at room temperature; DNase treatment can be performed concurrently. The lysis reaction is stopped, and the sample is then ready for reverse transcription or storage at –20°C for up to 5 months. Because samples are processed directly in culture plates (96- or 384-well), sample handling and the potential for sample loss or transfer error are minimized, resulting in higher reproducibility. Following sample preparation, a portion of the cell lysate is added to an RT reaction followed by real-time PCR using the included master mix.

Extraordinary Performance
To demonstrate the performance of the kit, replicate experiments were run using either traditional RNA purification and real-time PCR analysis, or the Power SYBR® Green Cells-to-CT™ Kit. The kit shows equivalent performance to results obtained using purified RNA for analysis of 10 to 100,000 cells. In addition, the sensitivity, efficiency, and dynamic range were shown to be superior to competitor lysate kits. Samples prepared with the kit have been tested for compatibility on various real-time PCR platforms including those from Applied Biosystems®, Roche, and Bio-Rad.

Accessory Product:
The SYBR® Green Cells-to-CT™ Control Kit (SKU# 4402959) contains a Xeno™ RNA control and primer set and is recommended to monitor the efficiency of cell lysis and amplification inhibition in Cells-to-CT™ reactions.
For Research Use Only. Not for use in diagnostic procedures.

Specifications

PCR Method: 2-Step qRT-PCR
Purity or Quality Grade: LD (Low DNA)
Reverse Transcriptase: M-MLV Reverse Transcriptase
Polymerase: AmpliTaq Gold® DNA Polymerase
Detection Method: SYBR
Primer-Probe Compatibility: Unlabeled Oligos
Reaction Speed: Standard
For Use With (Equipment): 7000 System, 7200 System, 7300 System, 7500 System, 7700 System, 7900HT System, GeneAmp® 5700, GeneAmp® 9600, StepOnePlus™, Standard Mode
Sample Type (General): Cells
Passive Reference Dye: ROX (Pre-mixed)
Green Features: Fewer resources used, Less hazardous, Less waste
Hot Start: Built-In Hot Start
Label or Dye: SYBR® Green
Product Size: 400 reactions
Shipping Condition: Dry Ice

Contents & storage

Store Stop Solution, Dnase I and 20X RT Enzyme Mix at -20°C. Store Lysis Solution, 2X SYBR® RT Buffer, PowerSYBR®Green PCR Master Mix at 4°C. Store Nuclease–free Water at room temperature

Documents

Manuals & protocols

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