Make the most out of your precious sample – With a lysate-based workflow there is no loss of RNA Thus, the ability to detect rare RNAs in low input cell samples (10–1,000) is higher compared to results from the same samples using RNA purification workflows.
Cells-to-CT kits measure relative gene expression by qRT-PCR without purify the RNA prior to amplification. A workflow that was designed in conjunction with real-time analysis to completely eliminate the need for traditional purification while maintaining performance relative to purified RNA. Featuring a unique method for lysing cultured cells while removing genomic DNA (gDNA) and preserving RNA integrity, the kits contain reverse transcription (RT) reagents for cDNA synthesis and available for both TaqMan® and SYBR® Green detection, enable real-time qRT-PCR analysis directly from cultured cells.
How the cells to CT kits work?
The lysis technology is designed for 10–100,000 cultured cells per sample. Cells are washed in PBS, then lysed 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 analysis on your real-time PCR instrument, or they can be stored at –20°C for up to 5 months.
Figure 1: The procedure for Fast Advanced Cells-to-CT kits is a 2-step process that involves reverse transcription and qPCR following cell lysis. A 1-step reaction is when RT and qPCR are performed in a single well. In the qPCR step, TaqMan Assays use custom primer and probe combinations designed for each target. These assays are more sensitive and specific than SYBR Green assays, which use generic qPCR primers.
Two formats available for increased flexibility
1. The TaqMan Fast Advanced Cells-to-CT Kit:
> 2 million predesigned assays- available for 28 species and some pathogens.
2. The SYBR Green Fast Advanced Cells-to-CT Kit:
Productive validated kit workflows complete sets of reagents preoptimized to work efficiently right out of the box.
A “Green” alternative to standard purification, environment in mind
True or False quiz:
- The product simultaneously isolate DNA and RNA, when leaving DNAse out of the reaction.
- I can use Heparin or EDTA blood.
- I can use the rapid lysate method for tissue samples as well.
- This method applicable for extraction of viral RNA.
- This method could be used for bacterial RNA extraction.
- I can use the direct lysis solution for uncultured primary cells or my other cells lines.