bad bad Agarose gel - (Jun/27/2009 )
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
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...
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
Rony on Jun 27 2009, 12:05 PM said:
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!!
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
claritylight
thanks alot i will try 4% and i will feed back
T C on Jun 27 2009, 10:11 PM said:
i did
Also, chk the grade of agarose, hope you are not using LMP agarose, which is fragile.
Best,
TC
LMP = low melting point.
Whatever agarose you're using is not behaving as expected. Can you post some details about it from the label?
Agarose will not gel if the pH of the buffer is acidic. Make sure your buffer is really a buffer.
Also, make sure that you boil your agarose gel thoroughly before casting it. Do let us know how it goes.