I cannot properly isolate sticky cell lines for the extraction of total RNA . I am getting low concentrations of RNA after extraction (poor bands on the agarose gel).
I guess you mean adherent cells. wait for the cells to be 90% confluent in a T75 flask, pour out media, wash in ice cold PBS, add '''very small'' amount of tyrpsin, allow cells to separate, deactivate trypsin, suck all material in a centrifuge vial, centrifuge at 6000rpm for 3 minutes, discard supernatant and then immerse the vials containing cells in liquid nitrogen. This will give you good amount of RNA
From this, the kit will give you good results though i used a different kit after going through this steps
If all you need is total RNA, I would advocate lysing the cells directly in the dish in a small volume of lysis reagent (e.g. Trizol) and scraping with a flat scraper or simply draining to a corner for collection. Do NOT add chloroform to the dish as it is likely to dissolve :-)
Trypsinisation (see excellent detailed protocol suggested by S. Karori above) also works, and is particularly important if you want to get FACS/protein/DNA/RNA/etc samples from the same dish. However, the inevitable delay in processing can cause reduction in RNA yield or integrity due to inherent (e.g. high RNase-content cell line) or logistical (e.g. inexperienced operator, overheating of sample etc) reasons.
Good luck!
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