The Luna Cell Ready Lysis Module has a lysis capacity of 10 - 100,000 cells in a 50 µl lysis reaction. Typically, 1-2 µl cell lysate (equivalent to RNA from 0.2 - 4,000 cells) can be transferred into a 20 µl downstream RT-qPCR. Most transcripts can be detected from 20 to 200 cells in a typical 20 µl reaction. However, some cell lines may contain high levels of inhibitors to cell lysis or RT-qPCR reactions. For a quick check of gene expression, ~1000 cells per 50 μl reaction is a good starting point for optimization. Please see additional FAQ’s for guidance regarding specific cell lines. Precious samples with few cells (≤10) can also be lysed with the Luna Cell Ready Lysis Module with subsequent detection using the Luna Universal Probe RT-qPCR Kit. However, modification of the Cell Lysis Mix is required as indicated below. Pilot experiments are recommended to optimize the entire workflow before using precious/limited samples.
| Components | Total volume (25 µL) |
|---|---|
| Luna Cell Ready Lysis Buffer (2X) | 12.5 µL |
| DNase I (RNase-free) (10X) | 2.5 µL |
| Luna Cell Ready RNA Protection Reagent | 2.5 µL |
| Luna Cell Ready Protease | 0.25 µL |
| Luna Cell Ready Stop Solution | 2.5 µL |
| Cells | Variable, up to 2.5 µL |
| RNase-free Water | To a final volume of 25 µL |