Invitrogen™ TaqMan™ Cells-to-CT™ Express Kit

Beschreibung

The TaqMan Cells-to-CT Express Kit is a complete cell lysate system designed for gene expression analysis directly from cultured cells without RNA purification. It enables RT-qPCR of the highest sensitivity and quickest workflow using just five components: Express Lysis Buffer, Express ezDNase, Express RT Master Mix, RT controls, and our best-in-class TaqMan Fast Advanced Master Mix. The TaqMan Cells-to-CT Express Kit provides the sensitivity you need in the fastest workflow available: RT-qPCR results for gene expression analysis in approximately 70 minutes.

Features of the TaqMan Cells-to-CT Express Kit include:

• Extraordinary ease and speed—96 samples ready for RT-qPCR in as little as 5 minutes
• No tedious RNA purification—no columns, heating, or centrifugation
• Verified workflows—complete set of reagents optimized to work efficiently right out of the box
• Environmentally friendly—the kit is REACH Compliant, has fewer components, and requires fewer pipetting steps reducing plastics costs
• Right sized kits for automation—available in 2500 reaction size that includes both lysis and RT components

Optimized chemistry with no need for stop solution

Gene expression analysis from direct cell lysis is made simple with our TaqMan Cells-to-CT Express Kit. With only one step for cell lysis and no need for the addition of a Stop Solution, our Express technology enables a rapid workflow with sensitive, accurate, and reproducible results. 10–100,000 cells are lysed using the Express Lysis Solution. The optional inclusion of ezDNase to the Express Lysis Solution enables removal of genomic DNA during cell lysis. Following a 5-minute incubation at room temperature, the lysate is used directly for reverse transcription using SuperScript IV VILO Master Mix. Once the RT reaction is complete, qPCR analysis is carried out using TaqMan Fast Advanced Master Mix and a TaqMan Gene Expression Assay.

Streamlined workflow allowing for rapid, accurate and sensitive results

RNA prepared using the TaqMan Cells-to-CT Express technology substantially expedites and streamlines gene expression analysis. Cell lysates exhibit sensitivity and specificity better than or equivalent to RNA extracted using traditional spin column isolation methods. The lysis procedure simultaneously prepares cell lysates for RT-qPCR and removes genomic DNA in only five minutes. The lysis step is simple to automate with robotic platforms for high-throughput processing of 96- or 384-well plates because it takes place entirely at room temperature. The procedure is also economical as there are only a few pipetting steps and, for cells grown in 96- or 384-well plates, no sample transfers. The high efficiency SuperScript IV VILO RT Master Mix transcribes even the most difficult (degraded and low abundant) RNA while reducing the typical reaction time by 10 minutes. Additionally, the TaqMan Cells-to-CT Express Kit is compatible with over 700,000 TaqMan Gene Expression Assays.

Maximize sensitivity of detection

Unlike some competitor kits that limit the amount of lysate in the RT reaction to 10%, the TaqMan Cells-to-CT Express 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.