Is autoclaving and sterilizing plastic benefit or harm RNA extraction?
I preferred to autoclave plastics before RNA extraction to assure they are clean especially they are from bulk bags, not high-grade ones.
On the other hand, some mention that autoclave water is full of RNAses!!
I was intending any way to snap freeze my samples directly in Liq. N2 and then store at -80 to inactivate RNAses
Thanks in advance
As you rightly say, autoclave water is not RNAse free, nevertheless RNAses will resist your autoclaving process if present in your plastic. Doing this you'll eventually disrupt DNases and other proteins and you'll kill bugs if present. This will not ensure the elimination of pyrogens such as LPS and also RNAses. I normally buy PCR-clean, RNAse/pyrogen free bags of vials and PCR-clean RNAse/pyrogen free pipette tips at least for RNA-related procedures. I always use this plastic with gloves previously treated with RNAse ZAP by ambion. I treat the surfaces where I extract with the same solution and periodically I also treat the RNA-extraction dedicated pipette sets with the same solution and the centrifuge rotor. I also never add my extraction reagents into a glass bottle, bottles even autoclaved are not RNAse free. I prefer 50 mL sterile tubes and then I periodically discard them for clean ones. Finally get DEPC water for your RNAs resuspension.
If you see someone gloveless with hands inside your RNAse free plastic bags then use a tanto knife for an emergency amputation (It's a joke but often this could happen, specially with thesis students).
As Paolo said, it would be better avoid autoclaved bags, tips and tubes and directly go for Nuclease-free plastic. I know that are more expensive but you will be sure that everything is done without external contamination.
Clean everything before to start with RNAse ZAP and add some RNA later to your samples before freezing.
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