C.B.S. Scientific GCMGU-202T - Manual (Page 13)

13 www.cbsscientific.com
SECTION 4
Running Conditions
4.1 Recommended Power
The recommended power conditions for optimal resolution are 50 to 150V, constant,
and 25 to 80 mA. The usual run time will vary for the voltage chosen but should
range from 15 to 60 minutes, monitoring nucleic acid migration by progress of marker
dyes. Constant power is not a necessity, but it produces uniform heat throughout the
run. Be sure the polarity is correct i.e. that the DNA is loaded near the cathode (black
electrode) to run toward the anode (red terminal).
Agarose gels may be stored for several days at 4°C wrapped in plastic wrap. Seakem
Agarose (FMC) is used (normally) for preparative and analytical gels. Other types of
agarose can be used for special purposes.
4.2 Recommended Buffers
Type Concentrated Stock/liter Final Concentration
TAE 50X - 242 gm Tris base 1X - 0.04M Tris-acetate
(Tris-acetate) 57.1ml glacial acetic acid 0.001M EDTA
100 ml 0.5M EDTA (pH 8.0)
TBE 5X - 54 gm Tris base 0.5X - 0.045M Tris-borate
(Tris-borate) 27.5 gm boric acid 0.001M EDTA
20 ml 0.5M EDTA (pH 8.0)
10X - Loading Buffer (DNA)
0.25% Bromophenol blue
0.25% Xylene cyanol
20 % Ficoll Type 400
0.1M EDTA, pH 8.0
4.3 References
1) Hames, B.D., Rickwoood, D. (ed.) (1990). Gel Electrophoresis of Nucleic Acids. A Practical Ap-
proach. 2nd edn. IRL Press, Oxford. Ch 2.
2) Sambrook, J., Fritsch, E.F., Maniatis, T. (1989). Molecular Cloning. A Laboratory Manual. 2nd
edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York. Ch 6.
3) Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., Struhl, K. (ed)
(1993). Current Protocols in Molecular Biology. Vol. 1, Greene Publishing Associates, Inc. and
John Wiley & Sons, Inc., Ch. 2.