Hi Everyone,
I am preparing some libraries from 150ng of mRNA. I am using a buffer to fragment my mRNA after isolation. I tried different concentrations of the buffer (from 0.2 to 1uL, also different times). I found out that 0.4 or 0.5 (5min at 70C) give me the best peaks. However, I think that I need to get bigger fragments. Should I target my fragmentation to get a peak over 200bp on bioanalyzer electropherogram or getting it around 170bp is okay? See picture attached.
Thank you.
I am preparing some libraries from 150ng of mRNA. I am using a buffer to fragment my mRNA after isolation. I tried different concentrations of the buffer (from 0.2 to 1uL, also different times). I found out that 0.4 or 0.5 (5min at 70C) give me the best peaks. However, I think that I need to get bigger fragments. Should I target my fragmentation to get a peak over 200bp on bioanalyzer electropherogram or getting it around 170bp is okay? See picture attached.
Thank you.
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