RNAse free water - Manufacturer (Nov/17/2005 )
Where can I get RNAse free water for cell culture...specifically for siRNA transfection.
I currently use Dharacom stuff...but was wondering if anyone else sells it.
Also can we use DEPC treated water for siRNA transfection??
I don't know where to buy it because I haven't done it, but I wanted to add a consideration...
Please make sure that your water is also endo-toxin free if you will add it to cells; DEPC water is not typically endotoxin free if you make it in the lab
Most companies that sell molecular biology products will sell nuclease-free water and it is cheap.
Eg: Invitrogen sells "UltraPure™ DEPC-Treated Water", cat# 750024, 4 x 100 mL for $43.00 USD.
I wouldn't use "home-made" DEPC-treated H2O for transfections since most labs do not treat and autoclave correctly.
I think you should get the nuclease free water from Ambion, it never sees DEPC so you don't ahve to worry about anything that would poison the cells....
I've used the nonDEPC treated nuclease free water from Ambion. I've ordered it in 50 ml bottles as well as the smaller individual vials.
Promega has the non-depc treated water..although they say that it is filtered and sterilized and very similar to what we would make in the lab. I am thinking of going with them since I can order it easily from the general sotres as opposed to making a new request which could take a while..also my advisor thinks it is a crime to pay 72$ for WATER....ambion..promega is 32 bucks
I already have my siRNAs in their buffer and rthis water is just for making the complexes just before adding it to the cells. So my questions are:
1. Can I get away with using just regular doubledistilled deionized sterile milipore water...
2. Once the transfection complex is made ie rna+trasfection agent...can RNAses still attack?? I think no..but what do u guys think..