Has anyone isolated tumor RNA from cell lines for use in cell culture? In all the papers I've read I can see no mention of isolating the RNA in any more sterile manner than is required for any RNA isolation. Does anyone isolate RNA for cell culture in the hood? I'm contemplating using the Roche High Pure RNA Isolation Kit as that seems very straight forward & I can do all the steps in the hood. Has anyone had experience using this kit for RNA isolation for cell culture? Thanks in advance for your input :)
Hi Melanie,
If you have sufficient cell numbers then why couldn't you use Trizol manual method inside the hood.
Hi Melanie!
I used to begin my isolation of RNA from melanoma cell lines in the hood using Qiagen RNeasy. I would make up fresh lysis buffer and then add it directly to my plated cells (in a 6 well plate). I would then leave for a short time room temperature, check under the microscope, and use a cell scraper to collect the material to one side of the well and pipette into an eppendorf tube. From there I would transfer to the bench top because I needed to use a mini centrifuge which we did not have in the cell culture room, but if you do then I'm sure all the steps could be performed in the hood!
By the way a side note, I found I would get varying amounts of RNA from different tumour lines even with the same input of cells.
Good luck with your RNA.
Thank you for the answers! We have both reagents so I'll give both a try & see how we get on :)
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