2. Gel Electrophoresis
Gel electrophoresis separates
molecules on the basis of their
charge and size. The charged
macromolecules migrate across a
span of gel because they are
placed in an electrical field. The
gel acts as a sieve to to retard the
passage of molecules according to
their size and shape.
3. One of the basic tools of modern
biotechnology is gene splicing.
This is the process of removing a
functional DNA fragment ( a gene) from
one organism and combining it with the
DNA of another organism to study how
the gene works.
The desired result is to have the new
organisms carry out the expression of the
gene that has been inserted.
4. The ability to cut and paste DNA
predictably is due to the use of restriction
enzymes.
They were first identified in and isolated
from the bacteria that use them as a
natural defense mechanism to cut up the
invading DNA of bacteriophages – viruses
that infect bacteria.
They are named for the
5. The negatively charged
particles move toward the
positive electrode while the the
positive charge particles move
toward the negative electrode.
6. electrophoresis work
• The gel is made from agar
• DNA is a negative molecules
• Molecules sort based on
•Charge
•Size
•shape
7. Agar comes from sea weed.
What is it used for?
The gel is 1% agarous and has
no electrical charge.
8. • DNA is cut into smaller fragments.
• Loading dye is used to indicate the
fragments of DNA are behind the dye
• The negative DNA molecule is
attracted to the positive electrode.
•The smallest fragments move the
greatest distance.
9. Remove comb and observe wells.
Place carbon paper in each end of the tray.
Cover with buffer, making sure the allow buffer to
overflow into each end of the tray.
Load gels.
Connect the electrodes.
Turn on power supply.
Allow gels to run – make sure you see bubbles
coming from the electrodes.
10. It will take about 30 minutes for the gel to
run.
Turn off power supply and remove
electrodes.
Pour off buffer into the designated
container.
Carefully remove gel from gel box and
place in glad container and cover with
stain.
Store in appropriate location.
11. They indicate that electrolysis of water is
taking place.
One electrode will have a lot of bubbles
and the other will have a lesser amount.
Why the difference?
The formula for water is H2O and the
splitting of the molecule will produce twice
as many atoms of hydrogen.
