4919396

Invitrogen™ TaqMan™ Gene Expression Cells-to-CT™ Kit

The TaqMan™ Gene Expression Cells-to-CT™ Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification.

Manufacturer: Fischer Scientific

The price for this product is unavailable. Please request a quote

Detection Method

Primer-probe

GC-Rich PCR Performance

High

Polymerase

DNA Polymerase

For Use With (Equipment)

7000 System, 7300 System, 7500 System, 7900HT System, Applied Biosystems StepOnePlus™ Fast Real-Time PCR System, StepOne™, Standard Mode, StepOnePlus™, Standard Mode, QuantStudio 6 and 7 Pro systems, QuantStudio 7 Flex system, QuantStudio 3 and 5 systems, QuantStudio 12k Flex system, QuantStudio 6 Flex system

Passive Reference Dye

ROX (Pre-mixed)

Product Type

Gene Expression Cells-to-C Kit

Reverse Transcriptase

M-MLV

Sample Type

Cell

Format

Tube

PCR Method

2-step RT-qPCR

Reaction Speed

Standard

Green Features

Fewer resources used and less waste, Less hazardous

Product Line

Ambion™, Cells-to-CT™, TaqMan™

Purity or Quality Grade

UP (Ultra Pure)

Shipping Condition

Dry Ice

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Description

  • The TaqMan™ Gene Expression Cells-to-C T ™ Kit makes it possible to perform expression analysis directly from cultured cells without RNA purification
  • This kit saves time and offers a simple workflow that is suitable for a few samples or can be easily incorporated into automated, high-throughput applications
  • The TaqMan™ Gene Expression Cells-to-C T Kit is: • Complete —optimized workflow includes cell lysis reagents with gDNA removal, RT enzyme mix, buffer, and new TaqMan™ Gene Expression Master Mix • Fast —7-minute sample prep, including DNase treatment, at room temperature • Easy —lyse samples in a tube or directly in culture plates • Robust —perform gene expression analysis on 10–100,000 cells per sample; results equivalent to those from purified RNA • Efficient —contains sufficient reagents to generate 500 real-time PCR results from 100 starting samples Featuring a unique method for lysing cultured cells while removing genomic DNA and preserving RNA integrity, the TaqMan™ Gene Expression Cells-to-C T ™ Kit contains reverse transcription (RT) reagents for cDNA synthesis and TaqMan™ Gene Expression Master Mix for real-time PCR analysis
  • TaqMan™ primer⁄probe sets are sold separately
  • Simple 7-minute sample preparation: part of a complete workflow Whether you are using plates or tubes, the TaqMan™ Gene Expression Cells-to-C T ™ Kit, which uses the simple 7-minute sample preparation procedure outlined (see figure), is designed for 10–100,000 cultured cells⁄sample
  • Cells are washed in PBS and lysed in solution for 5 minutes at room temperature; DNase treatment can be performed simultaneously
  • Lysis is terminated at room temperature by a 2-minute incubation with Stop Solution
  • The lysates are now ready for reverse transcription or storage 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
  • The TaqMan™ Gene Expression Cells-to-C T ™ Kit workflow enables unsurpassed gene expression evaluation with any of the >700,000 TaqMan™ Gene Expression Assays
  • This kit has been extensively tested for specificity with a broad selection of TaqMan™ Gene Expression Assays and shows performance equivalent to that obtained with purified RNA (see figure)
  • Achieve unsurpassed performance and sensitivity Unlike some competitor kits that limit the amount of lysate in the RT reaction to 5%, the TaqMan™ Gene Expression Cells-to-C T ™ Kit can accommodate 45% of the total RT reaction volume as cell lysate
  • Additionally, cDNA can comprise up to 45% of the real-time PCR reaction volume
  • The large lysate volume in the optimized RT reaction, along with the large cDNA volume in the subsequent real-time PCR using the TaqMan™ Gene Expression Master Mix, lead to maximum sensitivity
  • The master mix amplifies the target precisely and accurately, enabling the detection of small quantities of target, such as transcripts expressed at low levels
  • The ability to detect limited target quantities was tested by mixing a constant number of human cells with various numbers of mouse cells
  • The cell mixtures were prepared using the TaqMan™ Gene Expression Cells-to-C T ™ and assayed for a mouse-specific and human-specific gene
  • Comparative data were generated in the same manner with RNA purified by traditional methodology
  • The data show that RNA from as few as 10 mouse cells was detectable in a background of RNA from 10,000 human cells (see figure)
  • The ability of the TaqMan™ Gene Expression Cells-to-C T ™ Kit to detect relative low abundance transcripts was equivalent to that of purified RNA; at low levels, it was superior
  • Additionally, the performance of the TaqMan™ Gene Expression Cells-to-C T ™ Kit was compared to competitor lysate kits and to purified RNA
  • Inputs of 100–100,000 cells⁄lysis reaction were examined
  • The sensitivity of the TaqMan™ Cells-to-C T ™ Kit protocol was equivalent to that obtained with purified RNA, and it surpassed competitor lysates (see figure)
  • Furthermore, the lack of inhibition at high cellular inputs and the low variation among technical replicates demonstrate the reliability of this approach for gene expression studies using cultured cells
  • Proven performance, proven together All components of the TaqMan™ Gene Expression Cells-to-C T ™ Kit have been optimized for consistent and reliable performance
  • This removes the guesswork involved in assembling separate sample preparation, RT, and real-time PCR kits
  • And the TaqMan™ Gene Expression Cells-to-C T ™ Kit has been validated with TaqMan™ Gene Expression Assays for added quality assurance
  • Accessory product The TaqMan™ Cells-to-C T ™ Control Kit was designed for use with the TaqMan™ Gene Expression Cells-to-C T ™ Kit
  • 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
  • Order Info Shipping Condition: Dry ice

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Detection Method:
Primer-probe

GC-Rich PCR Performance:
High

Polymerase:
DNA Polymerase

For Use With (Equipment):
7000 System, 7300 System, 7500 System, 7900HT System, Applied Biosystems StepOnePlus™ Fast Real-Time PCR System, StepOne™, Standard Mode, StepOnePlus™, Standard Mode, QuantStudio 6 and 7 Pro systems, QuantStudio 7 Flex system, QuantStudio 3 and 5 systems, QuantStudio 12k Flex system, QuantStudio 6 Flex system

Passive Reference Dye:
ROX (Pre-mixed)

Product Type:
Gene Expression Cells-to-C Kit

Reverse Transcriptase:
M-MLV

Sample Type:
Cell

Format:
Tube

PCR Method:
2-step RT-qPCR

Reaction Speed:
Standard

Green Features:
Fewer resources used and less waste, Less hazardous

Product Line:
Ambion™, Cells-to-CT™, TaqMan™

Purity or Quality Grade:
UP (Ultra Pure)

Shipping Condition:
Dry Ice

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Detection Method:
TaqMan

GC-Rich PCR Performance:
High

Polymerase:
__

For Use With (Equipment):
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Passive Reference Dye:
__

Product Type:
1-Step qPCR kit

Reverse Transcriptase:
__

Sample Type:
Mammalian Cells

Format:
Tube

PCR Method:
1-step RT-qPCR

Reaction Speed:
7 min.

Green Features:
Fewer resources used and less waste, Less hazardous

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Cells-to-CT™, TaqMan™

Purity or Quality Grade:
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