CellAmp Direct TB Green RT-qPCR Kit

Brand	Takara Bio

The CellAmp Direct TB Green RT-qPCR Kit is designed for performing a simple two-step real-time RT-PCR, without RNA extraction, directly from various animal cells (e.g., adherent cells, floating cells, primary cells, various stem cells, and iPS cells). The kit makes it possible to prepare template samples from cultured cells and complete the analysis of reverse transcription reactions and gene expression in as little as 1.5 hr.

This product allows the effective elimination of genomic DNA. Therefore, it works well for analyses in which gDNA contamination is an issue, including situations in which the design of a primer pair spanning an exon junction is not possible or in which low-expression genes are to be analyzed. A two-step real-time RT-PCR reagent for TB Green detection is included in the kit, eliminating the need for reverse-transcription and PCR mixes to be prepared separately. This makes gene expression analysis easy to perform.

Overview

High-throughput analyses can be performed easily and rapidly (~1.5 hr)—there is no need to do RNA extraction or a heat-treatment to inactivate DNase, making the process faster and less cumbersome
Can be used with cells differentiated from stem cells and iPS cells—the improved lysis buffer works well with a variety of adherent cells, floating cells, primary cultured cells, various stem cells, iPS cells, etc.
Lysates can be stored for a long time—storage stability has been confirmed at –20°C for at least 6 months
Resistance to PCR inhibitors and high specificity—use of TB Green Fast qPCR Mix as the real-time PCR reagent makes this product highly resistant to PCR inhibitors and highly reactive for high GC-content targets

Applications

High-throughput gene expression analyses in a very short time (~1.5 hr)
RT-qPCR of stem cells (such as iPS cells, floating cells, adherent cells, primary cells, etc.)
Analysis of samples with high GC content-high tolerance to inhibitors and high specificity
Analysis of samples typically impacted by genomic DNA contamination

Gene expression profiling of iPSC-derived cardiomyocytes with CellAmp Direct TB Green RT-qPCR Kit

Gene expression profiling of iPSC-derived cardiomyocytes with CellAmp Direct TB Green RT-qPCR Kit. At 5, 18, and 42 days after the induction of cardiomyocyte differentiation, 50 µl of cell lysate was prepared, an RT reaction and a real-time PCR were performed, and a gene expression analysis was conducted. Purified total RNA was prepared as the control, and it was subjected to the same gene expression analysis. The gene expression levels at Days 18 (blue) and 42 (red) were compared with those at Day 5 (baseline). The gene expression profiles obtained from the lysates prepared with the kit were similar to that of the highly purified RNA.

CellAmp Direct TB Green RT-qPCR Kit

Additional information