Science Buddies: "Ask an Expert"

Hi all...
I have a query regarding the electrophoresis. I used 8% PAGE gel and i found that the dye+sample just stayed up in the wells. They didnt migrate down. What could be the problem ? I used 6X loading dye(2uL) and 6uL of sample.

Awaiting for some suggestions. Thanks in advance!

Hi,
It sounds like you might not have good electrical contact somewhere. What voltage and current are you using? Do you know that your power supply is working?

The other possibility is that the buffer that was used to make the gel was prepared incorrectly. There must be ions in the buffer to carry the electrical current.

Send more details about your gel prep and running conditions and we'll try to trouble-shoot your problem.

Sybee

Hi Sybee..
Thanks for your reply. My power supply is working earlier the same day i ran the gel. It was working. 1X TBE was used as running buffer.
Gel Prep: 10% PAGE with
30% acrylamide--4 mL,
10% APS--150 uL
TEMED- 9uL,
5X Tris--3.75 mL,
DI--7.1 mL.

Running conditions: 60 V for 90 mins.