Single Cell-to-CT™ qRT-PCR Kit
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Single Cell-to-CT™ qRT-PCR Kit
Invitrogen™

Single Cell-to-CT™ qRT-PCR Kit

Green features
The Ambion™ Single Cell-to-CT™ Kit contains a complete validated workflow for gene expression analysis for samples containing 1–10 cells. EachRead more
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Catalog NumberNo. of Reactions
445823750 Reactions
4458236400 Reactions
Catalog number 4458237
Price (USD)/ 50 reactions
1,392.65
Online exclusive
1,524.00 
Save 131.35 (9%)
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Add to cart
No. of Reactions:
50 Reactions
Price (USD)/ 50 reactions
1,392.65
Online exclusive
1,524.00 
Save 131.35 (9%)
Add to cart
Single Cell-to-CT™ qRT-PCR Kit
Catalog number4458237
Price (USD)/ 50 reactions
1,392.65
Online exclusive
1,524.00 
Save 131.35 (9%)
-
Add to cart

The Ambion™ Single Cell-to-CT™ Kit contains a complete validated workflow for gene expression analysis for samples containing 1–10 cells. Each kit contains reagents for sample preparation, reverse transcription, pre-amplification, and qPCR that have been optimized together in a simple workflow that can be completed in only 5 steps (see figure).

Key product features:

  • Pre-optimized workflow for real-time RT-PCR from single cells
  • Maximum sensitivity for single-cell analysis
  • Superior performance and reproducibility compared to alternative methods
  • Complete kit convenience and easy-to-follow protocol

Pre-optimized protocol ensures success and saves time

Each workflow begins with a simple 7-minute sample prep, where cells are effectively and reproducibly lysed with minimal processing into a solution that is compatible with downstream RT-PCR, without the need for purification. This is followed by the use of Superscript™ RT for reverse transcription, the TaqMan™ PreAmp Master Mix to amplify the cDNA, and TaqMan™ Gene Expression Master Mix for qPCR. All four of these steps have been developed to provide maximum sensitivity. The entire cell sample is maintained in the same well throughout the procedure so there is no sample loss or dilution of the precious limited material that could affect sensitivity.

Single cell sensitivity

Sensitivity of real-time PCR results can be affected by the efficiency of sample preparation, reverse transcription, or amplification. The Single Cell-to-CT™ Kit has addressed each of these potential problem areas to create a solution that enables reliable and robust gene expression analysis from single cells with maximal sensitivity. Detection of single cell equivalents using the Single Cell-to-CT™ Kit demonstrates appropriate linearity and sensitivity of qPCR results compared to a 100 cell sample control with excellent technical reproducibility (see figure). As expected, individual single cells exhibit greater variability due to inherent biological heterogeneity. In addition, through inclusion of a cDNA pre-amplification step, targets of interest are accurately amplified prior to real-time PCR. This critical step boosts signal in an unbiased manner, extending the analysis potential of limited samples. The accuracy and signal enhancement of a 96-gene panel using pre-amplification is shown (see figure).

Superior performance and reproducibility compared to alternative methods

Traditional sample preparation methods using organic solvents, glass fiber filters, or magnetic beads are not suitable for single-cell analysis due to the loss of sample through incomplete RNA precipitation, binding, and elution. Additionally, most single-cell homebrew methods which involve simple boiling to lyse cells lack the ability to inactivate endogenous RNases to stabilize gene expression profiles and can result in chemical cleavage of the RNA and inhibitor carryover, including gDNA contamination, which can affect all qRT-PCR applications. In contrast, the Single Cell-to-CT™ Kit shows superior sensitivity and reproducibility for single-cell analysis compared to traditional purification or homebrew boiling methods (see figure).

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Detection MethodTaqMan
PCR Method2-step RT-qPCR
Reaction Speed7 min.
For Use With (Equipment)7000 System, 7300 System, 7500 Fast System, 7900HT Fast System, 7900HT System, GeneAmp 9700, GeneAmp™ 2400, StepOne™, Fast Mode, StepOne™, Standard Mode, StepOnePlus™, Fast Mode, StepOnePlus™, Standard Mode, Veriti Thermal Cycler
Green FeaturesFewer resources used and less waste, less hazardous
Product LineAmbion™, Cells-to-CT™
Purification Time7 min.
Shipping ConditionDry Ice
For Use With (Application)RT-PCR
No. of Reactions50 Reactions
Sample TypeMammalian Cells
Starting Material1-10 Cells
Unit Size50 reactions
Contents & Storage
50 μL Single Cell DNase I, 0.5 mL Single Cell Lysis Solution (store at 4°C), 50 μL Single Cell Stop Solution, 75 μL Single Cell SuperScript™ RT, 5 mL TaqMan™ Gene Expression MasterMix (store at 4°C), 265 μL Single Cell Pre-Amp Mix, 150 μL Single Cell VILO™ RT Mix, Store kit components at -20°C except for Single Cell Stop Solution and TaqMan™ Gene Expression MasterMix, which should be stored at 4°C.