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How much loading dye? - (Oct/08/2008 )

I did a 10ul pcr, seperated the product into 2 5ul pcr tube.
1 of the 5ul tube, I have to digest it with my restriction enzyme
the other 5ul tube will go straight for gel electrophoresis.
Question is, how much loading dye(6x) must I add to the 5ul tube for gel running?

-Allson_1987-

QUOTE (Allson_1987 @ Oct 8 2008, 07:31 PM)
I did a 10ul pcr, seperated the product into 2 5ul pcr tube.
1 of the 5ul tube, I have to digest it with my restriction enzyme
the other 5ul tube will go straight for gel electrophoresis.
Question is, how much loading dye(6x) must I add to the 5ul tube for gel running?


1ul of loading dye (6x) + 5ul of sample

-Minnie Mouse-

so 2ul of loading dye if I were to run 10ul of sample ?

-Allson_1987-

QUOTE (Allson_1987 @ Oct 8 2008, 07:40 PM)
so 2ul of loading dye if I were to run 10ul of sample ?


yes

-Minnie Mouse-

i add 5ul every time when the sample size is 40ul, I add 10ul for anything greater than that.
I don't think it really matters if you add too much because it's just glycerol that makes the sample sink to the bottom of the well.

-airplane-