Under certain conditions mRNA may precipitate, particularly at high concentrations. Try heating the mRNA for 15 min at 37°C to improve solubility. Long incubations at elevated temperatures should be avoided.
Under certain conditions mRNA may precipitate, particularly at high concentrations. Try heating the mRNA for 15 min at 37°C to improve solubility. Long incubations at elevated temperatures should be avoided.
High concentrations of mRNA? 100ng/ul is considered high? and which buffer you may use to store mRNA, I had try RNA storage Solution from LT, and TE low EDTA (0.1mM) from USB and I’m about to try TE 1mMEDTA from Ambion. Still my mRNA activity drops after 3 months at -80C and stored at 100ng/ul. I’ll try to see if solubility may cause lower efficiency on the expression after some time on the freezer but concentration wise I haven’t seen a large difference. I’ll appreciate any help
Dear Marcia,
In general, 100 ng/uL is not considered a high concentration. We supply our mRNA at 1 mg/mL in 10 mM Tris-HCl, pH 7.5. That said, the mRNA sequence and structure can dictate its solubility. We recommend heating the mRNA for 15 min at 37°C to improve solubility. Long incubations at elevated temperatures should be avoided.
In regards to the diminishing activity of your mRNA, have you checked for degradation? In addition to the solubility, your issues with mRNA activity may also result from RNA degradation over time. To combat degradation you should use RNase-free reagents and materials and use proper technique. Additionally, we suggest that you aliquot your RNA to limit freeze/thaw cycles. A higher concentration may also improve stability however could exacerbate your issues with solubility.
Please let us know if we can help you further.
Regards,
Brea