Gels waiting for 90 min before transfer? - (Nov/19/2008 )
In our lab, we can do electrophoresis for 4 gels at one time but can only transfer 2 gels simultaneously due to limited transfer equipments.
If I do 4 gels, certainly 2 of them would have to wait in running buffer for more than one hour (one hour transfer plus manipulation time).
Would this ~90min idling of the gels lower the quality of the bands obtained in the end?
Thanks in advance!
I read the post "How to store PAGE gel and keep the bands after gel is run?". Seems that 90 min is nothing compared to the challenge over there (weeks). Probably ok to keep the gels in running buffer at 4 C for such a short time.
90 minutes is nothing. Think about the fact that you often have to transfer over a hour under constant voltage to pull the proteins out of the gel onto the membrane.
Probably all you'll do is make some of your bands a bit more diffuse.
Can you borrow a transfer apparatus from another lab?
We have the same set up here (i.e. our capacity for running gels is greater than our capacity to transfer to membranes). I either run two gels at a lower voltage so they finish later, or just remove the gel cassettes (the gel with the glass plates on either side) and put them in the fridge.
I find that bands diffusion occurs very quickly so if I leave a gel longer than 10 min before transferring things get soft. I would split up the run or annoy the PI to get another transfer rig. Gel rigs are cheaper than than time................
@bBop, They not the same thing, the electrical field is directional and the protein should stop once it hits the membrane.