Hello,
Your question is a bit vague. Are you trying to do patch-clamp/RT-PCR, get type-specific mRNA or total mRNA from all cells present in your culture (glias+neurons) ? Do you intend to do some kind of cell-sorting too? Good luck for your experiments
thanks for your kind answer thomas Launey above all fo your busyness actually i am tring to do RT-PCR first so thats why i am actualty interested to collect total mRNA from culture cell as this cell is different form other one that is my concern . is there any specific protocol for neuronal cell for isolating total mRNA ? and next i will try for patch-clamp as i am now learnnig this procedure its seems to me too much tough the patch-clam one ..........
For total RNA, standard method of extraction should work (actually easier from culture than from animal tissue, it is softer). How are you going to analyze the mRNA present in your sample? For PatchClamp/RT-PCR there are pioneer papers from the Rossier group, with all the method details. If you only need the RNA, no need to learn to patch (ie: record current), simply use a 3-5um pipette and aspirate the soma of your target. I hope this helps.
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