I know that DEPC treatment can make water RNase free. If I use DEPC-treated water for making PBS, I think, at the time of checking the pH the solution may be contaminated with RNase.
Is it possible to add DEPC directly into the PBS?
or please suggest any other suitable alternatives.
Thank you in advance.
It’s wise to buy DNAse and RNAse free water and prepare PBS buffer using that.
hi golam.. I am using milliQ water in both molecular and HPLC work, handle it carefully to prevent contamination or you can buy DNAse or RNAse free water commercially available in market. Nuclease free/ DEPC treated water are used for biological assays such as RNA, cDNA protocols. If you are using DEPC treated water then autoclave, filter and autoclave again. This will work great for purifying total RNA from tissue and making cDNA from the mRNA.
You can also buy RNAse out an add it to your buffer/samples. It's a strong RNAse inhibitor.
Thank you for your suggestions Rishi Sharma & Guadalupe Fernandez Milmanda
Hi Golam, in our Lab we also use milliQ water free RNAse to make buffer, how ever, RNAse also can exist on the buffer container or bottles
So, first we are rinsing our bottles with DEPC water, clean it up, and than used autoclave for 1 hour for further sterilization befor used as buffer container.
Hope it will help You
You can also buy pbs buffer rnase free, or add depc directly on the pbs buffer (1ml for 1L of pbs, and then autoclave 20 minutes)
Andi Muhammad Ichzan and Quentin Chartreux Thank you for your valuable comments.
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