Go Back   Science Forums Biology Forum Molecular Biology Forum Physics Chemistry Forum > Molecular Research Topics Forum > Molecular Biology Techniques > Agarose Gel Electrophoresis Forum
Register Search Today's Posts Mark Forums Read

Agarose Gel Electrophoresis Forum Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


Bromophenol blue question

Bromophenol blue question - Agarose Gel Electrophoresis Forum

Bromophenol blue question - Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


Reply
 
LinkBack Thread Tools Display Modes
  #1  
Old 09-20-2011, 06:57 PM
Pipette Filler
Points: 439, Level: 8 Points: 439, Level: 8 Points: 439, Level: 8
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Mar 2011
Posts: 8
Thanks: 0
Thanked 0 Times in 0 Posts
Default Bromophenol blue question



My bromophenol blue dye will not migrate across a gel reliably. Sometimes it seems to work properly but other times it will sit in the wells and refuse to move. The ladder and Gotaq Green commercial dye work fine on the same gel.

The recipe I use is

30mL glycerol
70mL ddH20
0.025g bromophenol blue

which I dilute to 1:6 with 1x TBE.

It will randomly work perfectly or fail completely to migrate into the gel. I have tried spinning it down and using the supernatant but that is still unreliable.
Reply With Quote
  #2  
Old 10-01-2011, 03:30 AM
butters's Avatar
M.D/Ph.D
Points: 4,938, Level: 47 Points: 4,938, Level: 47 Points: 4,938, Level: 47
Activity: 0% Activity: 0% Activity: 0%
 
Join Date: Dec 2008
Location: Malaysia
Posts: 315
Thanks: 25
Thanked 68 Times in 63 Posts
Default Re: Bromophenol blue question

Is the loading dye you make very blue?

I found it is ok to reduce bromophenol blue. Important regarding your question. Yes about 30% glycerol is 6x, so just add one part of you solution with five part PCR or DNA (yes you can add less).

If you dilute it with 1:6 buffer then your loading may not be able to bring all DNA to sit on bottom of well.

hmmmm, when you run, did you see bubble formation on anode or cathode? there are known cases where power pack was not connected/connected on the wrong power pack... check that out. If there are bubble mean voltage are apply.

and what voltage did you use for what length of gel?
Reply With Quote
Reply

Tags
blue , bromophenol , question


Thread Tools
Display Modes

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off
Trackbacks are On
Pingbacks are On
Refbacks are On

Forum Jump

Similar Threads
Thread Thread Starter Forum Replies Last Post
Serious question about white people and evolution.? Angeline Biology Forum 2 09-05-2010 05:06 PM
Raleigh Scattering - blue sky, red sunset Jim Coe Physics Forum 4 05-11-2005 08:34 PM
Question about blue light and red tinted glasses Christopher M. Physics Forum 6 01-31-2005 11:17 PM
Sci.chem FAQ - Part 1 of 7 Bruce Hamilton Chemistry Forum 0 01-15-2004 09:06 AM


All times are GMT. The time now is 02:00 AM.


Powered by vBulletin® Version 3.8.4
Copyright ©2000 - 2014, Jelsoft Enterprises Ltd.
Copyright 2005 - 2012 Molecular Station | All Rights Reserved
Page generated in 0.12372 seconds with 16 queries