Team:UNIPV-Pavia/Methods Materials/Electrophoresis

(Difference between revisions)

Revision as of 16:33, 22 September 2009

Protocols

(estimated time: 2 hours)

Prepare agarose gel in 1x TBE buffer
Add ethidium bromide (using gloves and face mask for your safety):
- 1 µl in the small size agarose gel (70 ml)
- 2 µl in the middle size agarose gel (150 ml)
- 4 µl in the big size agarose gel (250 ml)
Cast the gel, insert the well-forming comb and let it polymerize
Add the loading buffer to each sample
Load the samples and 8 µl of marker
Set to 70-100 volts and electrophorese for the required amount of time
Use UV-light to look at the bands
Take a picture of the gel, if needed (not when bands have to be cut!!!)

DNA samples

Ethidium bromide

Loading buffer 10x Blue Juice, Invitrogen

TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)
*54 gr Tris

*27.5 gr Borate

20 ml EDTA 0.5 M (pH 8)

Marker (1 kb DNA Ladder, Promega)

Face mask and gloves

Electrophoresis apparatus

Transilluminator

@@ Line 55: / Line 55: @@
 '''Loading buffer 10x Blue Juice, Invitrogen'''<hr color='white' width='70%'>
 '''TBE (Tris/Borate/EDTA buffer) 5x (final volume 1 liter)'''
-**'''54 gr Tris'''
+<br>*'''54 gr Tris'''
-**'''27.5 gr Borate'''
+<br>*'''27.5 gr Borate'''<hr color='white' width='70%'>
 '''20 ml EDTA 0.5 M (pH 8)'''<hr color='white' width='70%'>
 '''Marker (1 kb DNA Ladder, Promega)'''<hr color='white' width='70%'>