To get rid of microbial and related contaminations, buffers are autclaved after preparation. However, some buffers are not high temperature resistant such as MOPS, HEPES etc. Therefore, autoclaved water can be used for preparation of buffers. When the pH of the buffer is adjusted (buffer prepared in autoclaved water) will it have any negative effect(s) on enzyme assay or on other experiments? What happens if we use buffer prepared in non-autoclaved water.
Thats not a problem but the thing is you need to maintain the aseptic condition while weighing the chemicals and while adjusting the pH. i think its better to use membrane filters.
I agree with Rajesh,and if you care about the concentration of the constituent will be changed after filtration, you can make a 3X buffer for filtration and rinse the membrane with 2 volume of distill water.
You can do everything without filter sterilizing but you would also need to bake/ autoclave anything that comes in contact with the water when you make the buffer, or you can buy pre-sterile tubes/ containers. Or you can just make fresh buffer all the time and don't sterilize.
Hi, Thank you all for your time. I am not worried to get rid of microorganisms from my buffer. I expect not to use other protocols such as membrane filters or any other methods. The question is if autoclaved water will be used to prepare buffers for biological use, theoretically should any problem will be there or not?
Hello,
Certainly no effect on water
If you sterilize distilled or deionized water, then you can use it for preparation of buffer. However, by dissolving the salts, and setting of pH, it is again unsterilized.
What i suggest, is to prepare your buffer in non sterilized water, and then use membrane filter (generally known as Millipore) for sterilization.
Rasoul Shafiei so i want to know can be use uatoclave miliQ water for buffer preparation then filter it with membrane filter or first filter then autocalve. is there any difference?
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