Hello everyone,
Have anyone of you ever extracted both DNA and RNA from the PBMC sample stored in Qiazol!!! I know that we only can extract RNA from Qiazol but I also need the DNA from that sample. I found one Used developed protocol for that. I have attched it here. If anyone used that before, please suggest me or guide me if the protocol is fine to do both DNA and RNA extraction from Qiazol stored sample. If you have different idea on that, please share with me.
TIA.
I extracet RNA and DNA form cells and tissues using Trizol protocol. Trizol and Qiazol have similar composition (phenol/guanidine-based), but Qiazol is optimed for fat tissues. Follow the instruction of the datasheet that you found on Qiagen site wihtout problem
I have stored many times in Trizol, frozen, and months later isolated RNA, no problem. I have also stored in RNALAter and frozen and isolated RNA.
Thank you, Valeria De Arcangelis.
Just to make sure, you follwed the same guideline that I attached here, right! And also you got good DNA-RNA from triazol by following exactly the guideline without any modification!
Thank you Subhas C. Juneja. I also did RNA extraction from Qiazol. It was totally fine. But now I want to get DNA-RNA both, thats why I am looking for protocol that helps to extract both from Qiazol stored PBMC.
Hi, exactlu as you said I followed the dataseet guidlines. When you remove the trasparent supernatant to extract the RNA you don't touch the white interphase. Now you have to take also this interphase and follow the instruction to extract DNA.
I can only suggest you some things. In general I always leave few ul of transparent supernatant at the interphase, for precaution, to avoid contamination of RNA with DNA. If you want also the DNA remove all the supernatant that contin RNA, discard it and then recover the white interphase in another tube and procede to extraction.
You will extract RNA and DNA together so pay attention to the number of samples, remenber that you have to make a lot of centrifugations for two different protocols. Use two centrifuges if you can or organize centrifugation in correct way. Pay attention to contamination if you centrifuge all the sample together. Clean bench, glover and pipette, tips with filters. If you prefer start with RNA and, when you arrive to the last centrifugations, start with DNA extraction.
When you recover the white DNA interphase pay attention to the contamination with the protein under the interphase.
Good work
Valeria De Arcangelis , Thank you so much. Thats a great help. I will follow all of your suggestions.