Presentation
Collin College BIOL 1408 Unit 12 Biotechnology Packet
- Cours
- BIOL 1408
- Établissement
- Collin College
Collin College BIOL 1408 Unit 12 Biotechnology Packet
[Montrer plus]
Vendeur
S'abonner
Aperçu du contenu
Unit 12Biotechnology
This lab uses the following hazardous chemicals:
I. Ethanol
As a result students are required to wear, at minimum, Goggles & Gloves.
Abstract
DNA, or deoxyribonucleic acid, is the genetic material that is utilized by all living
organisms. Through a process called transcription, the information contained in the
sequence of DNA is copied into a strand of messenger RNA (mRNA). In eukaryotic
cells, the mRNA is altered during a process called splicing, and is then transported to
the ribosome. The ribosome “reads” the mRNA to create a specific polypeptide in a
process called translation. In this process, the ribosome, comprised of protein and
ribosomal RNA (rRNA), utilizes transfer RNAs (tRNA) that are specific for each amino
acid to construct the polypeptide chain. When translation is complete, this polypeptide
sequence folds into a functional protein.
This entire process of using DNA to encode mRNA to then encode a protein is referred to
as the “central dogma of biology.”
DNA ! RNA ! Protein
The proteins created by this process can be transported inside or outside of the cell to
perform their diverse array of functions. All proteins in our cells are originally made
through this process. Should some sort of mutation occur in the DNA, the resulting
protein may change, possibly resulting in a non-functional protein (see Section 12.4) or a
protein that performs its function too well or more often than is needed. This aberrant
activity can manifest itself via a variety of disorders and/or diseases, one of which is
cancer. In this lab you will utilize a variety of techniques intended to facilitate the
isolation of DNA as well as the analysis of this genetic material.
12.1 DNA Isolation
Introduction
Before analyzing the genetic material that is found at a crime scene, provided in a
paternity test, or extracted from an organism in the laboratory, one must ensure that the
sample in question is purified DNA. The techniques used to purify DNA are often times
referred to as DNA isolation techniques. These protocols are designed to facilitate the
156
,purification of DNA by separating the nucleic acids from lipids and proteins that are
found in the cell. Once isolated, the DNA may be used in a variety of experiments that
can qualitatively or quantitatively measure it. Such experiments will be further elucidated
in the later sections of this lab.
Materials and Methods
Steps 1- 5 will be done for the class as a whole. Each group will perform the experiment
from step 6 on.
1. Using a blender, blend half a banana in 250 mL of diH2O.
2. Pour the blended mixture into a 600 mL beaker and add 25 mL of Meat
Tenderizer solution and 25 mL of soap solution.
3. Use a fork to stir/mix the solution. Stir slowly so as not to create too much foam
from the soap.
4. Place the beaker on a hot plate until it boils, then remove it from the heat.
5. Line a plastic funnel with cheesecloth and strain the mixture through the
cheesecloth funnel into a new 400 or 600 mL beaker.
INDIVIDUAL LAB GROUPS START HERE:
6. Using a pipette and pump, aliquot 10 mL of the mixture into a large test tube
During the remainder of the experiment, you will be working with hazardous
substances. Goggles and Gloves must be worn from this point on!
7. GENTLY pour ice cold 95% ethanol down the side of the tube so that it makes
contact with the mixture. Fill the tube until it is 75% full, but be careful not to
disturb the banana mixture too much as the ethanol is poured.
8. Allow the test tube to sit, undisturbed, at room temperature for ten minutes. DNA
is not soluble in ethanol and will begin to come out of solution during this time
period. The DNA will appear as a cloudy mass with bubbles nestled in it.
9. A wooden splint or glass rod can be used to spool the DNA out o the ethanol
layer.
10. The tubes can be capped and kept as examples.
157
, Results
Please state what you observed when you added the Ethanol to the mixture, and if you
were able to remove the DNA from the tube, what it looked like.
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
_______________________________________________________________________.
12.2 Gel Electrophoresis
Introduction
Once DNA has been isolated, there are various techniques that can be utilized to analyze
different properties of the genetic material. Gel Electrophoresis is one such technique that
separates DNA molecules based upon their relative size.
Prior to running the DNA on a gel, DNA is cut in specific places using one or several
restriction enzymes. Restriction enzymes recognize specific sequences of nucleotides in
the DNA and clip the DNA into smaller pieces. Since our DNA is unique, DNA from
different individuals will be cut in different places, resulting in different sized DNA
fragments.
The cut DNA is then loaded into individual wells in the gel. The gelatin, known as
agarose, is a porous, semi-solid material, which allows molecules to move through it.
Smaller molecules will face less resistance when moving through the pores, and as such,
will run farther into the gel than larger molecules will in the same amount of time. To
ensure that the molecules remain in the gel and run in the same direction, an electrical
charge is placed through the gel. Since DNA has a negative charge associated with it, it
will run towards to positive end of the gel. The general set up of agarose gel
electrophoresis is pictured below:
158
Les avantages d'acheter des résumés chez Stuvia:
Qualité garantie par les avis des clients
Les clients de Stuvia ont évalués plus de 700 000 résumés. C'est comme ça que vous savez que vous achetez les meilleurs documents.
L’achat facile et rapide
Vous pouvez payer rapidement avec iDeal, carte de crédit ou Stuvia-crédit pour les résumés. Il n'y a pas d'adhésion nécessaire.
Focus sur l’essentiel
Vos camarades écrivent eux-mêmes les notes d’étude, c’est pourquoi les documents sont toujours fiables et à jour. Cela garantit que vous arrivez rapidement au coeur du matériel.
Foire aux questions
Qu'est-ce que j'obtiens en achetant ce document ?
Vous obtenez un PDF, disponible immédiatement après votre achat. Le document acheté est accessible à tout moment, n'importe où et indéfiniment via votre profil.
Garantie de remboursement : comment ça marche ?
Notre garantie de satisfaction garantit que vous trouverez toujours un document d'étude qui vous convient. Vous remplissez un formulaire et notre équipe du service client s'occupe du reste.
Auprès de qui est-ce que j'achète ce résumé ?
Stuvia est une place de marché. Alors, vous n'achetez donc pas ce document chez nous, mais auprès du vendeur danielhawkins1. Stuvia facilite les paiements au vendeur.
Est-ce que j'aurai un abonnement?
Non, vous n'achetez ce résumé que pour $5.29. Vous n'êtes lié à rien après votre achat.
Peut-on faire confiance à Stuvia ?
4.6 étoiles sur Google & Trustpilot (+1000 avis)
80364 résumés ont été vendus ces 30 derniers jours
Fondée en 2010, la référence pour acheter des résumés depuis déjà 14 ans
