C.B.S. Scientific MGU-303 - Manual (Page 9)

9 Mini-Fast Sub
SECTION 4
Running Conditions
4.1
Recommended Power
The recommended power conditions for optimal resolution are 50VDC / 25mA ≤ 60 minutes.
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
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
Ethidium Bromide Staining
One can add to the buffer only, the agarose gel solution only (after the solution has been
made molten and the temperature has dropped to 55°C), or can be added after the
electrophoresis run and the gel stained separately. Use at a final concentration of 0.1µ
g/ml from a 1mg/ml stock solution. Ethidium bromide is a Mutagen. ALWAYS wear
gloves. Dispose of solutions in accordance with the safety regulations of your institution.
4.3 References
1. Hames, B.D., Rickwoood, D. (ed.) (1990). Gel Electrophoresis of Nucleic Acids. A Practical
Approach. 2 nd edn. IRL Press, Oxford. Ch. 2.
2. Sambrook, J., Fritsch, E.F., Maniatis, T. (1989). Molecular Cloning. A Laboratory Manual. 2 nd
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.