 why gel is not running in straight line?? |
  |
 Nov 4 2009, 08:29 AM
Post
#1
|
|
|
member  Group: Active Members Posts: 11 Joined: 2-October 09 Member No.: 12885  |
Hello,
I have a problem with electrophoresis, my gels (regular cell lysates) are not running in a straight line, but thay are one side curved-up . does anyone have some idea why it is happening?? |
 |
 
|
|
Nov 4 2009, 08:39 AM
Post
#2
|
|
 Veteran Group: Active Members Posts: 747 Joined: 20-October 09 From: India!!! Member No.: 13357 |
smily effect...
heat is not uniform across the gel... try running at a lower voltage!!! wat conditions are u running in??? is it just the dye showing this effect or the bands too??? -------------------- Support bacteria - They are the only culture some people have!!!
Cheers!!! |
|
|
|
|
Nov 4 2009, 01:32 PM
Post
#3
|
|
 an elder Group: Active Members Posts: 555 Joined: 26-January 09 From: Staten Island, NY USA Member No.: 6470 |
dirty electrode.
-------------------- talent does what it can
genius does what it must i do what i get paid to do |
|
|
|
|
Nov 4 2009, 02:32 PM
Post
#4
|
|
 Veteran Group: Moderator Posts: 441 Joined: 11-September 04 Member No.: 4036 |
Tilted gel tray with different buffer depths at each side.
|
|
|
|
|
Nov 4 2009, 04:56 PM
Post
#5
|
|
 Veteran Group: Active Members Posts: 413 Joined: 26-January 09 From: Sydney, Australia Member No.: 6526 |
QUOTE (phage434 @ Nov 5 2009, 09:32 AM)  Tilted gel tray with different buffer depths at each side. or tilted gel tray when the gel is setting, which will lead to a gel of variable thickness. Do you have a picture? |
|
|
|
|
Nov 4 2009, 05:15 PM
Post
#6
|
|
|
Veteran Group: Active Members Posts: 213 Joined: 26-January 09 Member No.: 6449 |
It usually works ok for me when I decrease to a lower voltage.
|
|
|
|
|
Nov 4 2009, 06:56 PM
Post
#7
|
|
 Veteran Group: Active Members Posts: 115 Joined: 7-July 09 From: Missouri Member No.: 10808 |
How much protien lysate per well?
|
|
|
|
|
Nov 4 2009, 07:18 PM
Post
#8
|
|
 Enthusiast Group: Active Members Posts: 96 Joined: 6-August 09 From: Earth Member No.: 11584 |
this used to happen to me on the first and last well, I also cant figure it out why... maybe too high voltage is the culprit. My solution: i ignore the 1st and last well when i run my page gel...
|
|
|
|
|
Nov 5 2009, 12:03 AM
Post
#9
|
|
|
member Group: Active Members Posts: 11 Joined: 2-October 09 Member No.: 12885 |
Thanx everyone for your posts.
I usually run it at 70V at the beggining and 120-130 later, is it to high?? samples contain between 20-40 ug of protein in reduced condition. |
|
|
|
|
Nov 5 2009, 12:54 AM
Post
#10
|
|
|
Veteran Group: Active Members Posts: 747 Joined: 20-October 09 From: India!!! Member No.: 13357 |
QUOTE (porfirion @ Nov 5 2009, 01:33 PM) Thanx everyone for your posts. I usually run it at 70V at the beggining and 120-130 later, is it to high?? samples contain between 20-40 ug of protein in reduced condition. why dont u try running it at a constant voltage... instead of switching.. might help.. and 40 mcg is too much for a SDS-PAGE.. tat too iof u are silver staining it!!! -------------------- Support bacteria - They are the only culture some people have!!!
Cheers!!! |
|
|
|
|
Nov 5 2009, 09:06 AM
Post
#11
|
|
|
an elder Group: Active Members Posts: 555 Joined: 26-January 09 From: Staten Island, NY USA Member No.: 6470 |
QUOTE (adrian kohsf @ Nov 4 2009, 10:18 PM) this used to happen to me on the first and last well, I also cant figure it out why... maybe too high voltage is the culprit. My solution: i ignore the 1st and last well when i run my page gel... this happens because the edges run cooler than the central portion of the gel. -------------------- talent does what it can
genius does what it must i do what i get paid to do |
|
|
|
|
Nov 5 2009, 12:34 PM
Post
#12
|
|
|
Veteran Group: Active Members Posts: 175 Joined: 3-February 09 Member No.: 6937 |
The temp of the gel can make a difference. Do you circulate your running buffer? If the tank is small, you can stick it in an ice bucket and it will be fine. If it's large, you can set it on a magnetic stirrer with a stir bar underneath and run it at a lower voltage, or use an attachment for a circulating cold water bath.
-------------------- 42..."An immutable fixed-precision number of unlimited magnitude." http://en.wikipedia.org/wiki/Python_(programming_language), accessed 25June2009.
|
|
|
|
|
Nov 5 2009, 07:56 PM
Post
#13
|
|
|
Enthusiast Group: Active Members Posts: 96 Joined: 6-August 09 From: Earth Member No.: 11584 |
QUOTE (mdfenko @ Nov 6 2009, 01:06 AM) QUOTE (adrian kohsf @ Nov 4 2009, 10:18 PM) this used to happen to me on the first and last well, I also cant figure it out why... maybe too high voltage is the culprit. My solution: i ignore the 1st and last well when i run my page gel... this happens because the edges run cooler than the central portion of the gel. Oh....I didn't know that....haha... thank you for the enlightenment.  |
|
|
|
|
Nov 9 2009, 12:06 AM
Post
#14
|
|
|
Veteran Group: Active Members Posts: 213 Joined: 26-January 09 Member No.: 6449 |
Plus, avoid loading too much sample into one gel. You can always divide your sample volume into half and load them into two wells.
|
|
|
|
|
|
Lo-Fi Version | Time is now: 22 Nov 2009 - 09:24 AM |
| IP.Board Skin Developed By Creative Networks | ||
|
Home
- About
- Terms of Service
- Privacy
- Feedback
©1999-2009 Protocol Online, All rights reserved. |