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RNeasy Mini Handbook 10/2019
73
Appendix A: General Remarks on Handling RNA
Handling RNA
Ribonucleases (RNAses) are very stable and active enzymes that generally do not require
cofactors to function. Since RNAses are difficult to inactivate and even minute amounts are
sufficient to destroy RNA, do not use any plasticware or glassware without first eliminating
possible RNAse contamination. Great care should be taken to avoid inadvertently introducing
RNAses into the RNA sample during or after the purification procedure. In order to create and
maintain an RNAse-free environment, the following precautions must be taken during
pretreatment and use of disposable and nondisposable vessels and solutions while working
with RNA.
General handling
Proper microbiological, aseptic technique should always be used when working with RNA.
Hands and dust particles may carry bacteria and molds and are the most common sources of
RNAse contamination. Always wear latex or vinyl gloves while handling reagents and RNA
samples to prevent RNAse contamination from the surface of the skin or from dusty laboratory
equipment. Change gloves frequently and keep tubes closed whenever possible. Keep purified
RNA on ice when aliquots are pipetted for downstream applications.
Disposable plasticware
The use of sterile, disposable polypropylene tubes is recommended throughout the procedure.
These tubes are generally RNAse-free and do not require pretreatment to inactivate RNAses.
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