Hi,
I sterelized eppendorf tubes for RNA extraction samples by placing them in beaker covered with Aluminum foil and autoclave for 15 minute at 121°C.
Still there is little drops of water in the beaker. How can I dry it while keeping them sterile?
I thought to place them in an Oven at 65°C till dried.
Could the water if not dried affect the sterelity or the RNA extraction?
Thanks,
If you have purchased tubes that are not RNAse free, you will have to treat them properly before using. Only sterilization in autoclave will not destroy possible RNAses.
The protocol would be as follows:
1. Incubate tubes with 0.1% diethylpyrocarbonate (DEPC) treated water for 10-20 minutes, RT.
2. Autoclave the treated tubes to remove the DEPC. Not removing DEPC will strongly influence your downstream reactions.
3. Place the autoclaved tubes in the oven at 65*C and leave them to properly dry.
Good luck.
You must consider that autoclave not destroyed RNAse if you wanna to removing it.
I know autoclave will not remove RNAses but I wanted to clean the tubes as they are normal, not high-grade ones.
But why it is preferable not to autoclave? will it affect the RNA extraction process?
And that means that sterilization is not necessary for RNA extraction?
Thanks for your answers
Dear Mohamed, if your autoclave is equipped with a separate drying cycle program (30-40 minutes duration at least) then soon after the sterilization cycle finished just run this program a couple of times, the remaining drops of water inside the beaker or in eppendorfs will go off easily. Remember whenever there is moisture there is growth, so once you opened your sterile beaker to use the sterile eppendorfs and you find that there is still some moisture inside so it will be contaminated potentially.
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