2. What is 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. BIOTECHNOLOGY
♦ 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. Restriction Enzymes
♦ 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. How does electrophoresis work?
• The gel is made from agar
• DNA is a negative molecules
• Molecules sort based on
•Charge
•Size
•shape
7. What is agar?
Agar comes from sea weed.
What is it used for?
The gel is 1% agarous and has
no electrical charge.
8. How does it work?
• 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. Procedure
♦ 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. PROCEDURE (CONTINUED)
♦ 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. What is significant about the
bubbles?
♦ 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.
