Gel loading dye : sample ratio?? - have to follow strictly? (Feb/03/2010 )
hi, I'm new here.
I heard from a post-grad student that the loading dye-sample ratio have to be 1:5, if not the DNA wont run when running gel electrophorosis.
can some one clarify me? thanks.
P/S: I dont really follow the ratio, but i still get nice results.
the loading dye that the student gave you is a 6x (6 times concentrated) solution.
other formulations are 5x, 2x, etc. (although 6x appears to be most common for nucleic acid loading dyes).
for most consistent results you should prepare all of your samples the same way and knowing (and using) the concentration factor of the loading dye makes it easy.
You just need a bit to make the sample sink to the bottom of the well. Doesn't really matter how much IMO.
in our lab we used to put 3ul dye with 10ul PCR amplicon ,
mix them well and load the 10ul into the gel's well.
and this goes nice
microgirl on Feb 3 2010, 10:11 PM said:
yeah right, u dont have to be so precise....u just need to make sure that the sample settles down into the well....
i take it approximately...
The amount of loading dye will not influence your results, but you should be precise with the amount of sample if you need to analyse patterns from the gel. If you use changing amounts of sample you will not be able to compare your results!
I normally load 2ul of sample with 1 or 2 ul of 6X loading dye.
thank you all for the clarification