ThermoFisher Scientific

The mas ter mix delivers confident results in both single and duplex reactions, even in the presence of inhibitors commonly found in clinical samples. All lots are functionally tested to help ensure lot-to-lot reproducibility for Ct consistency and dynamic range across a wide variety of assays. This General Purpose Reagent is manufactured according to applicable CFR requirements and labeled For Laboratory Use. With stringent quality and premier performance, TaqPath qPCR Mas ter Mix is a superior choice for your diagnostic testing or development needs. Features of the TaqPath qPCR Mas ter Mix include: • Efficient and linear detection of up to eight logs with gene expression or miRNA assays • Enables reliable detection of low-copy templates with reproducible CT results • Robust multiplexing performance with exogenous or endogenous targets • Manufactured with stringent production and process controls to help ensure lot-to-lot consistency TaqPath qPCR Ma ster Mix, CG, is a 2X formulation designed for gene expression and miRNA analysis. It contains thermostable Fast DNA polymerase, uracil-N-glycosylase (UNG), dNTPs with dUTP, ROX dye (passive reference), and optimized buffer components for maximum robustness and reproducibility. Validated with a breadth of workflows TaqPath qPCR Mas ter Mix, CG, has been validated to provide high specificity and dynamic range for use in multiple real-time PCR applications. The formulation can be run in either fast or standard cycling conditions on a wide variety of qPCR platforms. The figure below demonstrates the excellent PCR linearity over an 8-log input range of template with both gene expression and miRNA assays. In addition, TaqPath qPCR Mas ter Mix, CG, has been engineered to retain consistent performance in preassembled reactions for up to 48 hours. The stability of this mix provides users of high-throughput, liquid handling systems the assurance that the results on the first plate will mimic those of the last plate. TaqPath qPCR Mas ter Mix has been benchmarked against similar competitor mas ter mixes and demonstrates equivalent or better sensitivity and dynamic range across a variety of targets. Reproducible sensitive detection We understand the importance of reliably detecting low-copy targets for your test quality and data interpretation. TaqPath qPCR Mas ter Mix, CG, helps generate significant and reproducible CT values for ≤10 copy detection. The figure below shows the consistent CT results obtained from three unique lots when detecting 10-copy inputs. This lot-to-lot CT consistency is preserved across multiple assays with different attributes and expression levels to maximize confidence in your results. Optimized for multiplexing Simultaneous amplification of multiple assays can be beneficial not only as a control to normalize and detect anomalies in experiments, but also to improve efficiency and cost-savings for labs. TaqPath qPCR Mas ter Mix has been optimized with enzymes and component concentrations to facilitate multiplexing while preserving specificity, and it is validated for duplex performance in each manufacturing lot. Inhibitor tolerant Unlike other mas ter mixes on the market, TaqPath qPCR Mas ter Mix’s unique proprietary formulation allows robust performance even in the presence of substances that can normally inhibit PCR, such as heparin, hematin, or EDTA, increasing your confidence when working with a variety of complex clinical samples. TaqPath mas ter mixes demonstrate increased tolerance to inhibitors compared to competitor mixes.

The TaqMan® Gene Expression Cells-to-CT™ 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-CT Kit is: • Complete—optimized workflow includes cell lysis reagents with gDNA removal, RT enzyme mix, buffer, and new TaqMan® Gene Expression Ma ster 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-CT™ Kit contains reverse transcription (RT) reagents for cDNA synthesis and TaqMan® Gene Expression Mas ter 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-CT™ 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-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 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-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® Gene Expression Mas ter Mix, lead to maximum sensitivity. The mas ter 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-CT™ 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-CT™ 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-CT™ 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-CT™ 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-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® Gene Expression Cells-to-CT™ Kit has been validated with TaqMan® Gene Expression Assays for added quality assurance. Accessory product The TaqMan® Cells-to-CT™ Control Kit was designed for use with the TaqMan® Gene Expression Cells-to-CT™ 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.

以最少的资源最大限度地提高处理通量。 在4QuantStudio™ 12K Flex系统上单次运行OpenArray®反应板，可将通量提高达32倍。 它相当于32个传统的384-孔qPCR板或者 12,000个数据点。 更多地提高你的吞吐量。 当QuantStudio™ 12K Flex系统配备有OpenArray®模块、9700 PCR仪和QuantStudio™ 12K Flex OpenArray® AccuFill™系统时，它可以在8小时工作日内生成110,000个甚至更多数据点。 对于需要最高通量的应用领域，添加QuantStudio™ 12K Flex自动化机器人和多个9700 PCR仪，您每天可获取逾2,000,000个数据点。 减少启动和操作时间 开始一个 12,000 数据点试验，时常往往需要不超过 20分钟。