Hoefer SQ33 Sequencer User Manual
Page 19
•
p8
3.4 Run Conditions and Buffer Volumes
1
Into the upper buffer chamber pour 800 ml
(approximately).
2
Into the bottom buffer chamber pour 800 ml
(approximately).
3
Ensure that there is no buffer leakage.
4
If desired pre-run the gel until the glass plates are
warm. Use the settings described below.
5
Prior to loading your samples flush out the wells with
running buffer to clear them of urea.
6
Heat the sequencing samples at 95 °C for 3 minutes,
place on ice and centrifuge 12,000 × g for 3 minutes.
Return to ice.
7
Load 1– 5 μl of sample using a suitable loading tip. If
possible avoid taking the sample from the bottom of
the tube — particulate materials may cause streaking
or smearing. Try to minimize sample dispersion, load
the sample directly onto the bottom of the well and
keep it as a thin layer.
8
Replace the safety lid ensuring it is positioned fully
down over the electrical connectors.
9
Connect and run the gel at the desired power setting.
The leads and electrical connectors are tested to
1,500 Volts and users are advised not to exceed
this voltage.
IMPORTANT! Do not fill over
the maximum fill lines.