gel drying problem - asking for help (Feb/18/2004 )
Hi,
I encountered a problem during drying gel for later phosphorimager scan. My gel is 15% PAGE/7M urea. After running I transferred the gel to Saran wrap, then covered with Whatmann 3MM filter paper, flipped over and put into gel dryer for drying. The temperature was set at 80 FC. The time was an hour. EVERYTIME when I stopped the vacuum and open the plastic cover of the gel dryer, my gel and Saran wrap over it were wrinkled and then my gel cracked. I am really frustrated. I would really appreciate it if anyone could tell me some trick or tip from your experience. Thanks in advance.
We never had such problem. We do it like this:
1) remove one panel of glass
2) apply filter paper directly to the gel
3) careful lift the fileter paper and the gel attaching to it
4) wrap the gel side of the paper with lab wrap and seal with tape at the edge of the other side
5) put one more paper on both sides
6) put the gel on the dryer with the side w/o wrap facing down
7) dry 1 hr at 80C
I hope this helps.
Thank you. I will try it.
6% acrylamide gels transfer to 3MM easily, 8% gels transfer about half the time, 10-20% gels are very difficult to transfer to paper. They can be transfered to GelBond from Cambrex, but this is expensive. The GelBond then replaces the Saran you are using and 2 pieces of 3MM paper are placed on the gel-side.
Only original SaranWrap is suitable for exposing wet gels, as all other (cling) brands are slightly porous and will let water contact the film, leaving black spots.
Wet the filter paper before placing it on the gel. It won't curl so much after drying and so you don't need to crack the gel in order to flatten it.
Hiya!
What's your gel thickness? In addition to the gel %, I am sure that >1mm gels are very difficult to dry without shattering.
Try a thinner gel.
Cheers!
Hi,
It seems you guys are not fixing the proteins in the gel before drying.
I do it by fixing the gel in normal fixing solution for 30 m and make sure to add the glycerol to fixer sol.
Hope this may help.
Niraj
You mentioned that you stopped the vacuum? Always wait until the machine is finished and there is no more suction....if you open it while there is still a slight vacuum seal, the gel will likely break.
When I dry gels for radiation work, I smooth the gel onto a piece of filter paper, and then loosely place the saran wrap over top. I place the plastic/rubber cover over it and smooth out any bubbles (as they can cause cracking too).
Also try putting the gel on a larger piece of filter paper...if it's just a bit bigger than the gel, it will probably curl and crack.
Good luck!
Katy on Sep 24 2009, 06:47 PM said:
if the gel is completely dry then you can just release the vacuum without fear of cracking. but, if it is even slightly wet then it will crack (or shatter) when you release the vacuum.