Description
The TaqMan Fast Advanced Cells-to-Ct Kit is a complete cell lysate system designed for gene expression analysis directly from cultured cells without RNA purification. TaqMan Fast Advanced Cells-to-Ct cell lysis reagents are supplied with highest sensitivity Fast Advanced reverse transcription enzyme mix, 2x RT buffer, and best-in-class TaqMan Fast Advanced master mix. The TaqMan Fast Advanced Cells-to-Ct Kit gives you all the sensitivity you need in the fastest workflow available: get gene expressions analysis results from an entire plate of cells in about 80 minutes.
• Complete—optimized workflow includes cell lysis reagents with gDNA removal, new Fast Advanced RT enzyme mix, buffer, and new TaqMan Fast Advanced 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
Unique method eliminates the need to isolate RNA from cultured cells
The Cells-to-Ct method features a unique lysis reagent that lyses cells and protects RNA. DNA removal occurs simultaneously in the 5-minute lysis step to save time and improve ease of use. Lysis is then stopped by the addition of a “Stop Solution” that irreversibly binds to the lysis reagents.
The TaqMan Fast Advanced Cells-to-CT Kit contains reverse transcription (RT) reagents for cDNA synthesis and TaqMan Fast Advanced Master Mix for real-time PCR analysis. TaqMan primer/probe sets are sold separately.
Simple 7-minute sample preparation
The TaqMan Fast Advanced Cells-to-CT Kit is designed for 10–100,000 cultured cells/sample. Cells are washed in PBS and lysed in solution for five 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 for a short term or -80°C for a long term.
Maximize sensitivity of detection
Unlike some competitor kits that limit the amount of lysate in the RT reaction to 10%, the TaqMan Fast Advanced Cells-to-CT 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 Fast Advanced 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.
Reduced sample loss and transfer error
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.
Compatible with gold standard TaqMan Gene Expression Assays
The TaqMan Fast Advanced Cells-to-CT 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.
Proven performance, proven together
All components of the TaqMan Fast Advanced Cells-to-CT 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 Fast Advanced Cells-to-CT Kit has been validated with TaqMan Gene Expression Assays for added quality assurance, giving you the sensitivity you need in the fastest sample-to-answer workflow available.