Guidelines for Working with RNA During RNA Cleanup
Successful RNA Cleanup requires using care when handling samples and ensuring the buffers and labware in contact with the RNA is free of contaminating nucleases.
To maximize RNA yield, integrity and purity, please keep the following principles in mind:
- RNases are stable and difficult to inactivate, and care must be
taken when handling samples during and after purification of RNA.
Plasticware and glassware in direct contact with RNA-containing
samples should be RNase-free. Gloves should be worn at all times
when handling samples and kit components. Frequent glove changes
are encouraged. Bench and equipment surfaces should be clean and
can be decontaminated prior to work using commercially available
cleaners such as RNaseZap®.
- Elution with nuclease-free water is standard, but for samples that
will be stored for use later, EDTA can be added to 0.1–1.0 mM
to limit degradation due to magnesium-requiring nucleases.
Alternatively, elution with slightly alkaline TE can be employed.
- Avoid unnecessary freeze-thaw cycles of purified RNA. Aliquots should be made, consistent with downstream needs.