Basically, I wanted to ask if we should prepare these components in RNAse free water and use directly or should we prepare them first and then autoclave them ? If we autoclave, wont it chnage the concentration a bit ?
Salts if heat stable (so possibly not some bicarbonates) and glycerol should be ok with autoclaving and freezing although phosphate buffers will often throw down a precipitate on freezing which may need warming to redissolve. If there are any detergents like SDS the freezing may form miscelles on freezing as the concentration increases during the freezing process which may need redissolving at raised temperature before using. Autoclaving should not change the concentration of salts if the storage vial is sealed so water cannot escape and so long as the material is mixed well to ensure any condensation on the inside of the lid/top of vial is redistributed back into the bulk of the buffer
You can treat the solutions with DEPC to destroy RNase activity, or use RNase-free water, RNase-free reagents, and baked glassware or RNase-free plastic containers.
See this useful web page:
https://bitesizebio.com/163/10-ways-to-work-rnase-free/
When autoclaving, you should not have an airtight seal because the container may explode. Use a tight foil cover. Autoclave screw-top bottle caps separately from the bottles, wrapped in foil. The amount of evaporation should be slight. Once the solutions have cooled, open the autoclave and replace the foil covers with the autoclaved caps. It's a good idea to use autoclave indicator tape to distinguish materials that have been autoclaved from those that haven't.
- Badges
- Science topic
- Similar topics
- Microbiology
- Assays