We want to extract RNA from small number of oocytes for RNA-seq. I am afraid that I lose RNA with column extractio. I am thinking about using Tirzol. Any idea?
hey, we used the Qiagen RNeasy Micro Kit for single bovine zygotes, and we had more than enough sample for RNAseq (but we did Cell-seq II, which needs little amount of initial sample).
Hi Pouya
I have used Qiagen Micro Kit too, and it worked well for 2 mg tissue. I could not extract good quality RNA from 2 mg tissue with the TRIzol method.
Hi!
I used Trizol and similar reagents many times to extract RNA from small samples (about 2 mg of tissue). Always cleaning at least twice with 75% ethanol before elution. After total RNA extraction, I usually purify the RNA with the RNEasy MinElute CleanUp (Qiagen) kit. I had some sample loss, but nothing to be afraid of and enough to make RNAseq. If you use this approach, clean the columns twice with the RPE buffer.
The other alternative would be to use the appropriate kit suggested by the colleagues above, or RNeasy Plus Micro Kit (Qiagen).
Best Regards!
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