Upon completion of DNA loading, the top is Gel is finally disposed in biohazard bag

placed back on electrophoresis chamber. and used TBE buffer in labelled beaker.

The black lead (-) was attached at the end

of the gel nearest the DNA.
The gel was visualized with Omega Fluor™
 The red lead (+) was attached at the Gel Documentation System and
other end. photographed.

Power supply was switched on.

 Voltage was set at 80 volts and gel was
Lid from electrophoresis chamber was
run for 45 min.
removed and gel was placed in container.

Once start button is pressed confirmation

of current flow is done by observation of After completion, power supply was
tiny bubbles rising from both the anode(+) turned off, the colours visible on gel was
and cathode(-). noted.
500μl buffer IR was then added
 Centrifuged at 12,00rpm for 1 minute
 Flow-through was discarded.

Using a NanoDrop™ 2000/2000c

The supernatant was discarded and
remaining pellet
700μl buffer WBwas
werecollected.
then added (ethanol
was previously added to buffer WB)
 Centrifuged at 12,00rpm for 1 minute
 Flow-through was discarded.
spectrophotometer, following values
were observed and recorded.
 Nucleic acid concentration
 The absorbance A260
 The ratio of A260/A280
The DNA was quantified and its purity
was analysed.
• The ratio of A260/A280

500μl buffer WB were then added

 Centrifuged at 12,00rpm for 1 minute
 Flow-through was discarded. 2 μl each of extracted DNA were subjected
to quantification and purity determination
 The rest of the samples were stored at 2-
8℃ for further use.
Spin-column AC was placed back to
Collection Tube
 Centrifuged at 13,00rpm for 1 minute
to remove ethanol residue in column

Flow through is elution solution containing

Spin-column AC was then placed into eluted DNA
a clean new centrifuge tube
 50μl buffer EB was then loaded in  The tubes containing DNA obtained were
the middle of the column. labelled S(A), T(B) and U(C)).
 Centrifuged at 12,00rpm for 1
minute