Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

bad bad Agarose gel


  • Please log in to reply
9 replies to this topic

#1 Rony

Rony

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 27 June 2009 - 06:46 AM

Hello everyone
am new here and i realy need help ASP
When I prepared Agarose gel 2% in TBE buffer I note that the gel disintegrate and rupture when i touch it with my finger tp place it in the electrophoresis

what to do with this gel i cant buy new agaros i already have alot of it what to do ???


can any one help :)


Regards

#2 HomeBrew

HomeBrew

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 930 posts
15
Good

Posted 27 June 2009 - 07:06 AM

Welcome to the BioForums, Rony!

A 2% agarose gel in anything should be quite sturdy... Are you sure you didn't accidently make a 0.2% gel? A 2% gel should have 2 grams of agarose per 100 ml of buffer...

#3 Rony

Rony

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 27 June 2009 - 09:05 AM

hi

thanks for wellcoming me HomeBrew:D

yes i add 2gram in 100ml

befor the gell was ok until the old agarose end so i use another one i found in the lab and i face this proplem :(

is there any thing i can add to solidfy this bad agarose in prepration

:)

#4 claritylight

claritylight

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 104 posts
0
Neutral

Posted 27 June 2009 - 01:49 PM

hi

thanks for wellcoming me HomeBrew:D

yes i add 2gram in 100ml

befor the gell was ok until the old agarose end so i use another one i found in the lab and i face this proplem :(

is there any thing i can add to solidfy this bad agarose in prepration

:)


you let your 2% gel completely dry correct? i set mine to dry for 30mins as a standard and don't touch it. then i transfer it to the electrophoresis

you can try making a higher percent gel and see what is going on, like a 4%. if the 4% still is bad, i'd buy some new agarose!!

#5 T C

T C

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 277 posts
0
Neutral

Posted 27 June 2009 - 09:11 PM

Or chk the balance if it is calibrated and you are actually weighing 2g and not less.

Also, chk the grade of agarose, hope you are not using LMP agarose, which is fragile.

Best,
TC

#6 Rony

Rony

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 28 June 2009 - 03:01 AM

claritylight

thanks alot i will try 4% and i will feed back

:) :lol: :lol:

#7 Rony

Rony

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 28 June 2009 - 03:08 AM

Or chk the balance if it is calibrated and you are actually weighing 2g and not less.

i did :lol:

Also, chk the grade of agarose, hope you are not using LMP agarose, which is fragile.

what u mean by LMP agarose??

Best,

TC



Thanks TC :)

#8 HomeBrew

HomeBrew

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 930 posts
15
Good

Posted 28 June 2009 - 05:04 AM

LMP = low melting point.

Whatever agarose you're using is not behaving as expected. Can you post some details about it from the label?

#9 phage434

phage434

    Veteran

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,268 posts
213
Excellent

Posted 28 June 2009 - 12:45 PM

Agarose will not gel if the pH of the buffer is acidic. Make sure your buffer is really a buffer.

#10 jiajia1987

jiajia1987

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 212 posts
0
Neutral

Posted 28 June 2009 - 10:24 PM

Also, make sure that you boil your agarose gel thoroughly before casting it. Do let us know how it goes.




Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.