7
E. Gel Pouring
1.
Fit the casting dams over each end of the tray and place onto a level
surface. The dams should be fitted so that there is no gap between the
sides of the tray and the groove in the dams. This will ensure that there
is no possibility of gel leakage.
2.
Place the gel casting tray on a flat surface or use the Roth levelling
table (Art. No. N854.1).
3.
Insert the appropriate comb into the grooves. Melt agarose in
electrophoresis buffer.
Cool the agarose down to 50-60 °C in order to
avoid leaking of the gel during pouring and any damage to the gel
tray.
Pour the agarose to the desired height (approx. 5 mm).
4.
Do not move the casting tray until the gel has polymerised. We
recommend further polymerisation of the agarose gel for approx. 10 min in the refrigerator. Place
the gel casting tray and agarose gel into the electrophoresis chamber and submerse the gel in
running buffer.
Note: We recommend using 0.5x TBE buffer for optimal signal-to-noise ratio in blue light transmission.
F. Performing real-time nucleic acid separation
1.
With runVIEW placed on an even bench surface, switch it on using the ON/OFF button located at
the rear of the base unit.
2.
Place the gel tray containing an agarose gel in the middle of the electrophoresis tank in the correct
orientation (the wells in which samples are to be loaded should be closer to the black/negative
electrode)
3.
Pour in enough of your electrophoresis buffer so that the gel is just submerged.
4.
Load the DNA samples. We recommend use with the DNA loading and staining reagents
ROTI
®
Load DNAstain1, 2 or 3 containing the non-toxic fluorescent dye EvaGreen
®
(see below).
5.
Select your settings accordingly. In order to run the system at constant voltage, switch the mode
button to the Voltage setting and alter the value to the desired setting as described in
Set Up Mode
(the Volt LED will be illuminated by this stage).
6.
Use the same principle to run the system at constant current (in this case the Current LED will be
illuminated instead).
7.
For separations free of condensation, connect the cable from the runVIEW lid into the rear of the
base to activate the extractor fan (IMPORTANT
– see Note, next page).
Note: 1.) In order to operate under constant voltage or constant current modes, adjust the other
parameter to the maximum value. For example, to operate under constant voltage, adjust the current
to the maximum output of 300mA before running the power supply with the voltage set at the desired
output setting.
For constant runs, “time” should be set to zero. “Time” then counts up until the
Start/Stop-button is pressed.
2.) We recommend using 0.5x TBE buffer for optimal signal-to-noise ratio in blue light transmission.
To Start the Run
1.
Press the
button to commence electrophoresis. Press the
button again to pause or
stop electrophoresis at any time.
2.
Press the
button to switch on the blue light source in order to view real-time DNA migration.
In order to conserve the blue light lamp, the light will be there for 10 seconds only and switch off
again by its own. Press the button for 3 seconds to switch constant light on.
3.
Once electrophoresis is completed ‘End’ will show in the display accompanied by an alarm.
Press the
button again to cancel this.
