How to preserve southern membranes - (Jan/08/2006 )
Hi,
After a failed labeling of a probe I took the already pre-hybed membranes and washed them in 6XSSC and kept them for 2 days in the same solution. Usually I know that they're kept in 0.5XSSC and 0.1XSDS. Is the higher concentration of salts a big mistake? Are my membranes ruined?
Thanks... 
at which temperature did you keep your membranes?
when i want to preserve membranes, i wrap them in saran as they are still wet and store at -20°
when i want to preserve membranes, i wrap them in saran as they are still wet and store at -20°
it was RT. I took them out of there after 2 days and replaced the solution with 0.5SSC and SDS, but do you know if something bad might have happened to the bounded DNA?
Did you bake your membrane after transfering with SSC? If you did that, DNA has already bounded the membrane very well. Then after failure of hybridization, you just need to wash away the probe, (should be in formamid something, you can refer to Molecular Cloning) and wrap it with saran and keep it at -20C for whenever you want.
Yes, the membranes are baked (actually this would have been the 3'rd probing, and the last one) and I had them prehybridizing at 42 deg when I had to abort since the labeling failed. So all I did was wash the Hybrisol out of them with 6Xssc and leave them inthere. I just want to know if 6XSSC can do anything funny to well-prepared membranes if kept for 2 days at room temp...
hi
i think your membrane is still ok.
stripping should be more abrasive than leave in 6Xssc at RT for 2 days...
Thanks to all of you... next time I'll refrain from daydreaming when using buffers. 