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Versión/Version 1, Sept 2017
5.
Pull the comb(s) out carefully and move the tray with gel to the main tank.
7.
Running the gel
1.
Mix the sample with loading buffer (see point
6. Solutions
).
2.
Pour buffer (TAE or TBE) into the tank till the gel is immersed, almost 1 mm higher. Thus
will complete the experiment in shorter time and better quality of sample resolution.
3.
Load the samples into the wells with pipette. Multi-channel pipettes can be used with MC
compatible combs to load samples.
4.
Carefully cover the tank with lid and connect it with a power supply.
5.
Typically gels run under 90V- 50V. Be noted that, generally higher voltage enables faster
electrophoresis but poorer quality of sample resolution.
6.
Run electrophoresis.
8.
Gel Staining and Viewing
1.
Put the gel containing the appropriate volume of 0.5ug/ml ethidium bromide to a
staining box and stain for 15~30 minutes, see solutions (P11)for stock stain
concentration and adjust to the volume used accordingly. The staining box should be
covered.
Note:
Ethidium bromide is a suspected carcinogen and the necessary safety
precautions should be undertaken.
2.
De-stain the gel for 10~30 minutes in distilled water again ensuring the gel is
completely immersed.
3.
Rinse the gel twice for a couple of seconds with distilled water.
4.
Put the gel in a UV Transilluminator.
5.
The samples will often appear as brighter, clearer bands when photographed or
viewed using a gel documentation system. However if the gel bands are too faint then
the staining procedure should be adjusted so that there is less de-staining. If there is
too much background then the staining procedure should be adjusted so that there is
more de-staining.
9.
Buffer solutions
1x TAE
40mM tris (PH 7.6), 20 mM acetic acid, 1 mM EDTA.
To prepare stock of TAE 50X (1L) dissolve in 750 ml of distilled water:
•
242 g tris base (FW=121)
•
57.1 mL glacial acetic acid
