I am extracting RNA from 10%DMSO+FBS, cryopreserved PBMCs. I am performing RNA seq. Although I am getting decent RNA yield, it has come to my attention that the DMSO may affect the RNA quality. Because we have limited sample remaining, I would prefer to minimize washes if possible. I was going to fast thaw and dilute the sample before pelleting , but not do a PBS/HBSS was prior. your thoughts/ expert advice?
Thanks!
You are getting Decent RNA yield. I doubt DMSO will affect adversely on RNA quality. DMSO will be washed away during centrifugations while you are isolating RNA, anyway.
I recommend using Qiagen RNeasy plus MIni kit or Qiagen RNeasy mini kit ( does't have the gDNA eliminator spin column) but it is good too and If you care about DMSO it will get washed during your RNA purification. Make sure you see the quality of your RNA using nanodrop and see the reading of A260/280 and 260/230.
Please see this paper for further information regarding the impact of DMSO on the transcriptomeArticle RNA-Stabilized Whole Blood Samples but Not Peripheral Blood ...
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