Agarose gel powder ( FMC/BioWhittaker). Choose the agarose according to the purpose as below:
Purpose
Agarose for DNA fragment > 1k bp
Agarose for DNA fragment < 1k bp
Analysis
SeaKem ME
Nusieve 3:1
Preparative
SeaKem GTG
Nusieve 3:1
Gel apparatus
Erlenmeyer flask
Stir bar
MilliQ water
Microwave oven
Pipette
Procedure
Weigh the amount of agarose according to the following table, and put in Erlenmeyer flask. Add 50 ml (or 49ml) H2O. Put a
magnetic stir bar in the flask. Cover a small flask on the mouth of the big flask and microwave until the gel melt (1min + 10").
% Agarose (w/v)
Size Fragments Separated (bp)
0.5
1,000 to 30,000
0.7
800 to 12,000
1.0
500 to 10,000
1.2
400 to 7,000
1.5
200 to 3,000
3-4
10 to 1000
Cool (in 60oC water bath) and add 1 ml 50X Electrophoresis Buffer and 0.5ug/ml ethidium bromide, mix.
Pour gel into gel aperatus (put comb at proper position). let solidify at room temperature. Take off the comb and flush wells. Add 1x electrophoresis buffer to cover the gel about 1mm higher. The gel is ready for loading samples.