Help on probe striping from Northen blot

dinakar bhattramakki ddh at unlinfo.unl.edu
Thu Jun 22 12:52:51 EST 1995


biolc4 at jetson.uh.edu wrote:
: Hi netter,

: I have some difficulty in stripping the probes from my Northern blot.
: I did something like to treat the filter with SSC and SDS at 95 C about 
: 20 minutes, all RNA is gone.  I would really appreaciate if somebody 
: can tell me an effective but a little mild method to do this kind of
: work.

: Thank you for your attention.

: biolc4 at jetson.uh.edu 

I use the following procedure to strip the membrane of radioactivity 
before reprobing. I have successfully reprobed upto 6 times without 
any problem. (MSI Nylon membranes)

1. Stripping membrane: Incubate 10 m at RT
     0.1 N NaOH + 0.2 % SDS
2. Neutralize 10 min at RT
     0.2 M Tris, pH 7.5 + 0.1 X SSC + 0.2 % SDS
Good luck.   -Dinakar

--
Dinakar Bhattramakki/Ph.:(402) 472-6243/email:ddh at unlinfo.unl.edu



More information about the Methods mailing list