Hello everybody.
Does anyone know if I could freeze my infected THP-1 macrophages before extracting RNA?
The reason to do so would be that I would like to perform controls to check if the cell culture would be contaminated during the process.
In my head it would be like this: Day 1) Infect Cells and seed samples to verify contamination--> 24h 37ºC; Day 2)Take samples from each well infected and seed in plates to evaluate if there were any contaminants. While I wait for the controls to positive or not, freeze the plate in -80ºC.
Day x) If the controls do not positive in ~10days continue with RNA extraction and reverse transcription.
PS: I have looked about RNA Protect Bacteria Reagent (qiagen) to aid in RNA stabilization. Does anyone has experience with this product?
I routinely freeze the cells in Aurum RNA lysis buffer, which is basically guanidinium thiocyanate, beta-mercaptoethanol, etc., and store in -20C until i feel like extracting RNA after months and up to maybe a year or two (got one study still frozen). I used this on HeLa's, SUPT1s, and (PBMCs) and see no reason why it wouldn't work for anything else.
Hello all.
Thank you very much for your help in this matter. It surely will make our experiments much easier and make the results much more reliable.
Best regards.
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