I would be very grateful if the community could give me some tips: as these organisms are very small, I wonder how many polyps do I have to pool for one RNA extraction or if one is enough? What is the best lysis protocol? How many RNA were you able to extract and quantify with NanoDrop before the RT-PCR?
António Maximiano Fernandes
I agree with Alim. The critical point is during extraction from tissue. Trizol is good method. If 100 mg is not enough try to do optimization. Blinder equipment may improve the extraction. During centrifugation make sure that debries deposit on bottom to avoid interferente. In nanodrop 100-200 nanogram (ng) is ok
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Yashwant Chavan
Trizol is the best method. You just need to take care that tissue is completely chopped into cells so that you get better yield.
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