I was wondering what kind of quantities of total RNA have people used for extracting mRNA from? At our lower limits, we have 5ug, max usually 10ug. Is this a common amount to be using with success?
Also, having had trouble with the purification of mRNA, we are now looking at amplifying it to get the quantity we need. Has anyone done this with success from the quantities of total RNA above? Any comment on the potential bias introduced with the PCR and selection steps in terms of NGS results would be appreciated.
Thanks!
Also, having had trouble with the purification of mRNA, we are now looking at amplifying it to get the quantity we need. Has anyone done this with success from the quantities of total RNA above? Any comment on the potential bias introduced with the PCR and selection steps in terms of NGS results would be appreciated.
Thanks!
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