Everytime when I seed HD11 at 2*10^6 cells/mL into 6-well plates overnight, and after treated cells by reuterin about 6h, I can't extract cellular RNA, even the untreated cells. I have tried to use Trizol and centrifugal adsorption column but all failed. I am depressed and can't find the solution, please anybody help me...
Could you give us a little bit more information about the problem you are having? The following info would really help.
- How did you collect the cells from the plates?
- How much Trizol did you use and did you follow the standard protocol exactly?
- Which column kit did you try, how much lysis buffer did you use, and did you follow the standard protocol exactly?
- How much water/buffer did you elute your RNA in after extraction?
- When you say 'failed' what exactly do you mean? No RNA, or dirty samples (bad ratios), or degraded RNA, or qPCR failure, or something else? What method/s did you use to check whether your RNA extraction worked?
Sorry to answer your question with more questions but I need to narrow down what your problem is in order to help. And sorry to hear that you feel depressed about it! Molecular biology can be a big learning curve but it does get easier and I'm sure your problem is solvable with some thinking and testing :)
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