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

Catalog No.	No. of Reactions
44-582-36	400 Reactions
44-582-37	50 Reactions

Catalog No. 44-582-36 Supplier Invitrogen™ Supplier No. 4458236

Description

The Ambion Single Cell-to-C_T™ Kit contains a complete validated workflow for gene expression analysis for samples containing 1–10 cells.

The Ambion Single Cell-to-C_T™ 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 five steps.

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-C_T 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-C_T Kit demonstrates appropriate linearity and sensitivity of qPCR results compared to a 100 cell sample control with excellent technical reproducibility. 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.

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-C_T Kit shows superior sensitivity and reproducibility for single-cell analysis compared to traditional purification or homebrew boiling methods.

• 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
• Suitable for use with 1-step and 2-step qRT-PCR
• Less hazardous, less waste, fewer resources used

Order Info

Shipping Condition: Dry ice

Specifications

• Content And Storage: 400 μL Single Cell DNase I, 4.0 mL Single Cell Lysis Solution (store at 4°C), 400 μL Single Cell Stop Solution, 600 μL Single Cell SuperScript™ RT, 50 mL TaqMan™ Gene Expression MasterMix (store at 4°C), 2.1 mL Single Cell Pre-Amp Mix, 1.2 mL 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.
• Detection Method: TaqMan
• PCR Method: 2-step RT-qPCR
• Reaction Speed: 7 min.
• For Use With (Equipment): 7000 System, 7300 System, 7500 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
• Product Line: Ambion, Cells-to-CT
• Purification Time: 7 min.
• Quantity: 400 reactions
• Shipping Condition: Dry Ice
• For Use With (Application): RT-PCR
• No. of Reactions: 400 Reactions
• Sample Type: Mammalian Cells
• Starting Material: 1-10 Cells

For Research Use Only. Not for use in diagnostic procedures.