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leen.dimashkieh
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FACULTY OF HEALTH SCIENCES

DEPARTMENT OF MEDICAL LABORATORY TECHNOLOGY

Students’ Names:
Leen Dimashkieh 202202737
Lama Mardini 202202925
Samia Zaatarieh 202200926

Genetics and Molecular Biology– MELS401


Section 1

Keywords: Bands, DNA, gel, electrophoresis, migration

1
Contents of electrophoresis buffer & it’s uses:
We used TBE in this experiment:
However, TBE and TAE buffers are typical electrophoresis buffer
solutions in molecular biology.
Gel electrophoresis is a typical technique in molecular biology and
biochemistry for separating and analysing molecules, such as
DNA, RNA, and proteins, depending on their size and charge.
Electrophoresis buffer is a fluid used in this procedure. In the
electrophoresis process, the buffer fulfils a number of crucial
functions.

1. Ionic Conductivity: The electrophoresis buffer's main job is to


supply ions—typically borate and chloride ions—that make it
easier for current to pass across the gel. This permits charged
molecules to flow through the gel while the electric field is acting
upon them.

2. pH Control: To keep the gel's pH at a particular level,


electrophoresis buffer is utilised. The pH of the buffer can be
changed to provide ideal circumstances for the separation of
particular kinds of molecules, such proteins or DNA.

3. Safety: By avoiding excessive heat accumulation and preserving


constant temperatures, the buffer guarantees the safety of the
electrophoresis process.

2
Contents of loading dye & why we use it:
It contains glycerol, methylene blue, and buffer. Methylene blue
helps in visualizing the sample as it migrates through the gel by
coloring it. Glycerol is mixed with the sample to make it denser
than the electrophoresis buffer which helps the sample sink into the
walls at the top of the gel.
Interpret electrophoresis migration results:

1 2

Migration of 2 samples is shown on the gel, which migrated


depending on the size of the band (the smaller the band the faster
the migration) and the width of the band indicate the concentration
of the molecule. For further interpretation we compare the results
with the reference ladder.

Briefly list the steps:


Agar preparation:
1. Dilute concentrated 50X Electrophoresis buffer with distilled
water.
2. Mix agarose powder with buffer solution in a 250ml flask.

3
3. Dissolve agarose powder by boiling the solution. Microwave
the solution on high for 1 minute. Carefully remove the flask
from the microwave and mix by swirling the flask. Continue
to heat the solution in 15-second bursts until the agarose is
completely dissolved (the solution should be clear
like water).
4. Cool agarose to 60C with careful swirling.
5. While agarose is cooling, add 4 uL of Ethidium bromide dye.
6. Pour the cooled agarose solution into the prepared gel-casting
tray. The gel should thoroughly solidify within 20 minutes.
7. Remove end caps and comb.
Running the gel:
1. Place the gel (still on the tray) into the electrophoresis
chamber. Cover the gel with 1X Electrophoresis Buffer.
2. Place safety cover on the unit. Check that the gel is properly
oriented.
3. Connect leads to the power source and PERFORM
electrophoresis. Allow the tracking dye to migrate at least
3.5 cm from the wells.
4. After electrophoresis is complete, remove the gel and casting
tray from the electrophoresis chamber.
5. Transfer the gel to the UV Lamp.
6. Visualize results using a white light visualization system.

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