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Tips for Working with RNA
When removing tissue or cells from the source (animal, tissue culture plates, etc.) all materials that come into contact with material should be sterile -
RNA degrades quickly!
Gloves
should be sterile and/or strayed with RNAse Decontamination spray or 70% EtOH before use and between samples
Avoid touching objects that contain RNAses (face, clothing, benchtop)-if this does occur, switch gloves before proceeding
All
tools
should be autoclaved and avoid touching unnecessary objects when not in use
This includes tissue culture plates, surgical tools, tubes for sample storage, etc.
Tools should be sterilized between samples by submerging in EtOH briefly
All
buffers
(PBS, tissue culture medium, etc.) should be cold and filtered before use
Keep cold in between uses
Wearing a face mask is also recommended before isolation procedure
Samples should be snap-frozen in liquid nitrogen if not immediately processed in tubes that are sterile
Keep material for RNA isolation cold AT ALL TIMES
During isolation procedure, tubes should be kept on ice or at 4°C
A refrigerated microcentrifuge is recommended
Use filter-tip pipettes that are sterile
After isolation, extracted RNA should be kept on ice or stored at -80°C
Never allow RNA to thaw on benchtop, always on ice!