Resuspension of Oligos and siRNAs
Note that our siRNAs are delivered annealed and in solution at a concentration of 40 uM. If you have dried down siRNAs for longterm storage you should follow the protocol described below to resuspend your siRNAs.
What is the best way to resuspend siRNAs?
- Do a short spin to collect the pellet at the bottom of the tube
- Add appropriate amount of provided RNAse-free water
- Heat 5 minutes at 37°C and mix
- Use siRNA (freeze after use)
Alternatively:
to disrupt higher aggregates, which may have formed during lyophilization and to increase the efficiency of your siRNA you may:
- Do a short spin to collect the pellet at the bottom of the tube
- Add provided RNAse-free water to a final concentration of 40 uM
- Heat 2-3 minutes at 90°C
- Let siRNA slowly cool down to room temperature
- Use siRNA (freeze after use)
What is the best way to resuspend single strand RNA Oligos?
- Do a short spin at max. speed in a centrifuge to collect the pellets at the bottom of the tubes.
- Add the appropriate amount of sterile water or buffer (RNase-free!).
- Heat 2 mins. at 65 °C.
- Vortex or mix by pipetting vigorously up and down.
What is the best solvent for an Oligo?
For all Oligos (except those with fluorescent dyes) sterile RNase-free water or buffer (e. g. 10 mM Tris-HCl pH 7.5) is fine. We recommend dissolving fluorescently labelled Oligos in 10 mM Tris pH 7.5 as degradation may occure at low pH or high pH (distilled water may have a pH of 6.0!). However Cy-labelled Oligos (Cy3, Cy3.5, Cy5, Cy5.5) degrade slowly at pH >7.5 and should therefore be dissolved at pH 6.0 -7.0. Note that certain dyes are very sensitive to oxidation.
