5-2
Horizontal System
Thermo Scientific
Section 5
Technical Tips
Chart A
Mobility range of DNA in different percentage agarose gels
Agarose % (w/v) . . . .Approximate range of separated DNA fragments (kb)
0.3 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .60 to 5
0.5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .30 to 1
0.7 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .12 to 0.8
1.0 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .10 to 0.5
1.2 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .7 to 0.3
1.5 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .4 to 0.2
2.0 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .3 to 0.1
3.0 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .<0.1
It should be noted, an increased agarose % gives better separation of small
fragments and also bands very close together that tend to be more difficult
to separate, visualize and photograph. A specialty agarose product
formulated to increase resolution of low molecular mass samples may also
be used.
Example:
A good mid-range gel percentage would be 0.7%, or 0.7g.
agarose in 100mls. electrophoresis buffer (TBE or TAE), following heating
and dissolving the agarose, 10ul of ethidium bromide stock solution
(5mg/ml) is added. The gel would be run with compatible electrophoretic
running buffer (1X TBE or 1X TAE) that also contained ethidium
bromide. 1 liter of the running buffer would contain 100ul of this 5mg/ml
ethidium bromide stock solution.
Reagent Information
