Any one using "innuPREP DNA/RNA Mini Kit" from Analytik Jena fro RNA isolation? I have a problem, tried isolating RNA from SH-SY5Y cells but failing at the first step itself. Not getting any pellet after lysis with RL solution provided in the kit. Some one please help.
I was struggling with extraction of RNA from iPS cells but finally I used Rnasy mini plus kit Qiagen I adjusted the number of cells with the RL plus solution such as (1 well (6-well plate) with 80% confluence ) treated with 170 ml RL plus solution after collecting the cells from the well and I used Qiashredder tube before adding 70% ethanol
With SH-5y cells growing as 2 subsets of cells, one of them being loosely adherent, are you starting with an adequate cell # for lysis? We have great success using the Cells to CT kit from Ambion (Life Tech). It is an in-well lysis and collection, with the lysis mix able to be stored at -20C for several weeks before running the RT and qPCR.
Also, just basic Trizol method works excellent, as well.
The problem is, even before I joined my lab they bought this "innuPREP DNA/RNA Mini Kit". According to the protocol its very easy. I am planning to repeat the isolation process once more with more number of cells. Thanks a lot all for your answers.
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