Bio-Rad Fish DNA Barcoding Kit - Manual (Page 9)

Fish DNA Barcoding Quick Guide 7
Quick Guide
Lesson 3: Gel Electrophoresis
1. Retrieve the 5 µl samples of PCR products (4
samples) from 4°C. To each one, add 5 µl of
sterile water. Use a new pipet tip each time.
2. Add 2 µl of UView ™ 6x loading dye to each
sample, using a new pipet tip each time. Mix
samples well and pulse-spin.
3. Set up your gel electrophoresis apparatus as
instructed.
4. Load the agarose gel in the following lane order
and volumes, using a new pipet tip each time:
Lane Sample
1 – EMPTY
2 – EMPTY
3 – 20 µl PCR molecular weight ruler
4 – 12 µl (+) E
5 – 12 µl (–) E
6 – 12 µl 1 E
7 – 12 µl 2 E
8 – EMPTY
5. Ask your instructor whether the electrophoresis
buffer your electrophoresis units contain is 0.25x
TAE or 1x TAE.
If your buffer is 0.25 x TAE, run the gel at 200 V
for 20 min.
If your buffer is 1x TAE, run the gel at 100 V for
30 min.
6. Visualize the gel on a UV transilluminator or
imaging system. No gel staining is required
as the loading dye contains a fluorescent
compound that will allow visualization of DNA
with UV light.
5 µl
sterile water
1, E 2, E (+), E (–), E
2 µl
6x loading dye
1, E 2, E (+), E (–), E