We are trying to make a library for RNA-seq. We used magnetic beads to purify RNA, but the concentration of RNA after purification is not good.
We used NON-salt wash buffer to wash the beads because proteins are generally soluble in a low-ion strength solution (see the following URL). However, I saw a lot of researchers recommend to use HIGH-salt wash buffer to wash the beads?
I am grateful if somebody tell me the reason why high-salt wash buffer is recommended although proteins do not solve well in this condition.
Thank you very much for your help!
https://www.tankonyvtar.hu/en/tartalom/tamop425/0011_1A_Proteinbiotech_en_book/ch01.html
Which kind of magnetic beads are you using to purify RNA? Agencourt?
I usually start extracting RNA with Trizol followed by chloroform extraction and alcohol precipitation first to get rid of the majority of proteins and only in later stages of the protocol I purify the RNA (now dissolved in RNA-free water) with Agencourt RNA Clean XP beads.
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