How to get rid of radioactive probe from nylon membrane? - (May/16/2006 )
Do you know, how to get rid off the radioactive probe from nylon membrane to use it again but with different probe?
How to make sure, that no probe left from previous experiment?
I would go to the protocol portion of this website and look up the membrane-stripping protocols; there should be a number of options from which to choose
Because there are a number of options online, I am asking which one do you mostly recommend, simply which is the best...
I have used Ms Colitti's method from the protocol page (use a hood, that stuff is stinky....P U)
except I add a modification...I do the treatment step at about 50C, per Pierce's advice
others may have different methods?
i prepare boiling 0.5% SDS solution add it over the membrane and let all come back to RT under a hood.
So...you just boil 0.5% SDS and put the membrane in it and that 's it? How you make sure, that you don't have any leftovers of previously applied radioactive probe on the membrane?
i just boil the SDS solution and put it on membrane. Yes. I put the stuff on a platform rocker, agitate gently til the solution is at RT.
I do 2methods for detecting residual radioactivity : for 32P, i use a counter device, or an instant imager. For 35S i use instant imager.
You may try then 1%SDS if you have too many radioactivity remaining.
Thanks again for your great advice.
I will use this method for my purposes.
What is more I use TRIzol for RNA isolation from protoplasts and it works great! Also, I'm dissolving my samples in 100% formamide to prevent degradation.
Thank you very much :-D