Bio-Rad PV92 PCR Informatics Kit - Manual (Page 5)

Lesson 3 Electrophoresis of Amplified
PCR Samples and Staining of Agarose
Gels
1. Obtain your PCR tube from the thermal cycler
and place in the capless micro test tube.
Pulse-spin the tube for ~3 seconds at 2,000 x g.
2. Add 10 µl of PV92 XC loading dye into your
PCR tube and mix gently.
3. Place an agarose gel in the electrophoresis
apparatus. Check that the wells of the agarose
gels are near the black (–) electrode and the
base of the gel is near the red (+) electrode.
4. Fill the electrophoresis chamber and cover the
gel with 1x TAE buffer. This will require ~275 ml
of 1x buffer.
5. Using a clean tip for each sample, load the
samples into 8 wells of the gel in the following
order:
6. Secure the lid on the gel box. The lid will attach
to the base in only one orientation: red to red
and black to black. Connect the electrical leads
to the power supply.
7. Turn on the power supply and electrophorese
your samples at 100 V for 30 minutes.
Staining of Agarose Gels
1. When electrophoresis is complete, turn off the
power and remove the lid from the gel box.
Carefully remove the gel tray and the gel from
the gel box. Be careful, the gel is very
slippery. Nudge the gel off the gel tray with
your thumb and carefully slide it into your
plastic staining tray.
37
+
–
+
–
Centrifuge
Lane Sample Load Volume
1 MMR (DNA standard) 10 µl
2 Homozygous (+/+) control 10 µl
3 Homozygous (–/–) control 10 µl
4 Heterozygous (+/–) control 10 µl
5 Student 1 20 µl
6 Student 2 20 µl
7 Student 3 20 µl
8 Student 4 20 µl