biochem lab final exam
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Apr 3, 2024
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Mass (g) = Molarity (M, mol/L) x Volume (L) x Molecular Weight (g/mol)
C
1
V
1
= C
2
V
2 // M
1
V
1
= M
2
V
2
Concentration (g/L) = Molarity (mol/L) x Molecular Weight (g/mol)
LAB 2: Inducing Taq Expression
1.
How will you monitor the growth of bacterial culture in Lab 2?
a.
By plating bacteria on agar plates and counting the colonies
b.
By using a pH meter
c.
By using a microscope with 10,000x magnification
d.
By using a spectrophotometer 2.
What reagent will you use to induce protein expression in Lab 2?
a.
LB media
b.
Lysozyme
c.
IPTG
d.
Ampicillin
3.
In what phase of bacterial growth will you harvest the cells?
a.
Lag phase
b.
Exponential (log) phase
c.
Stationary phase
d.
Death phase
4.
In what phase of bacterial growth will you induce Taq DNA Polymerase?
a.
Lag phase
b.
Exponential (log) phase
c.
Stationary phase
d.
Death phase
5.
What plasmid is used in Lab 2 for protein expression? a.
pBR322
b.
pAKTaq
c.
pAMP
d.
pKan
6.
What is the purpose of adding IPTG to LB media in Lab 2?
a.
To help bacteria grow faster
b.
To prevent growth of unwanted bacteria
c.
To induce protein expression
d.
To stop all the bacteria from growing before collecting them
7.
What is the purpose of adding ampicillin to LB media in Lab 2?
a.
To help bacteria grow faster
b.
To present growth of unwanted bacteria
c.
To induce protein expression
d.
To stop all the bacteria from growing before collecting them
8.
What molecule does IPTG bind to?
a.
It binds lactose
b.
It binds the Lac repressor
c.
It binds the beta-lactamase
d.
It binds the lac promoter
9.
What molecule does ampicillin bind to?
a.
It binds lactose
b.
It binds the Lac repressor
c.
It binds beta-lactamase
d.
It binds the lac promoter
10. What method will be used to collect bacteria from the culture after desired protein is expressed?
a.
Pelleting bacteria from centrifugation
b.
Collecting bacteria on a filter
c.
Plating bacteria on agar plates with ampicillin
d.
Bacteria don’t need to be collected
Lab 3: Purification via Heat Precipitation
1.
What is the function of lysozyme?
a.
It solubilizes the lipids in the cell membrane
b.
It digests the peptidoglycans (in the cell wall of bacteria)
c.
It prevents protein degradation
d.
It digests bacterial DNA
e.
It protects DNA from DNAse
2.
Why will you heat your sample to 75°C during Lab 3?
a.
To denature E. coli DNA
b.
To degrade E. coli DNA c.
To denature E. coli proteins
d.
To degrade E. coli proteins
e.
To denature Taq DNA polymerase
3.
How will you separate contaminating proteins from Taq DNA polymerase?
a.
By using lysozyme
b.
By using protein chromatography
c.
By using salt extraction
d.
By using heat
e.
By using a specific filter
4.
Why do you need to add PMSF to your samples during Lab 3?
a.
It solubilizes the lipids in the cell membrane
b.
It digests peptidoglycans
c.
It digests bacterial DNA
d.
It prevents protein degradation
e.
It is used to denature E. coli proteins
5.
How is Taq DNA polymerase different from E. coli proteins?
a.
It has a different heat tolerance
b.
It has a different size
c.
It has a different charge
d.
It contains a special signal sequence absent in E. coli
e.
It contains amino acids that are not found in E. coli
6.
What method will be used to separate the bulk of E. coli proteins from Taq DNA polymerase at the end of Lab 3?
a.
Lysozyme treatment
b.
DNAse treatment
c.
Salt precipitation
d.
Restriction enzyme digest
e.
Centrifugation
7.
What method will be used to separate denatured E. coli proteins from Taq DNA polymerase sample at the end of Lab 3?
a.
Chromatography
b.
Filtration
c.
Differential centrifugation
d.
Salt precipitation
e.
Restriction enzyme digest
8.
At the end of Lab 3, Taq DNA polymerase will be found in what fraction?
a.
Pellet
b.
Supernatant
c.
Equal amounts in the pellet and supernatant
d.
Mostly the pellet, but some in the supernatant
9.
Differential centrifugation separates cell contents based on
a.
Charge
b.
Amino acid sequence
c.
Nucleotide sequence
d.
Density
e.
Shape
10. Which of the following is not a component of the cell wall of E. coli?
a.
Outer membrane
b.
Peptidoglycans
c.
Inner membrane
d.
LPS
e.
They are all components of the cell wall
Lab 4: Purification via PEI Precipitation & Dialysis
1.
When pH > pI of the protein, the overall charge of the protein is
a.
Negative
b.
Positive
c.
Neutral
d.
Depends only on salt concentration
e.
You can’t tell
pH > pI = negative
pH < pI = positive
pH = pI = neutral
2.
What fact is NOT true about PEI?
a.
It is a negatively charged polymer due to many amino groups
b.
It binds to many proteins in solution based on charge
c.
It binds to DNA and RNA
d.
It precipitates Taq DNA polymerase out of solution\
e.
It is a polyelectrolyte
3.
What is the overall charge of Taq DNA polymerase at pH 7.9?
a.
Neutral
b.
Positive
c.
Negative
d.
It depends on the salt concentration
e.
You can't tell
4.
How will you separate Tag DNA polymerase from impurities after adding PEI to your samples?
a.
By filtration
b.
By heating
c.
By salting out
d.
By centrifugation
5.
In what fraction will Tag DNA polymerase be after PEI precipitation?
a.
Pellet
b.
Supernatant
c.
Equal amounts in the pellet and supernatant
d.
Some in the pellet, but mostly in the supernatant
6.
What is the purpose of the salt wash step?
a.
To salt in Taq DNA polymerase
b.
To salt in contaminating proteins
c.
To salt out Taq DNA polymerase
d.
To salt out contaminating proteins
7.
Why is dialysis necessary at the end of Lab 4?
a.
To remove contaminating protein from Taq
b.
To increase Taq concentration
c.
To decrease Taq concentration
d.
To increase salt concentration in the sample
e.
To decrease salt concentration in the sample
Lab 5: Purification by Chromatography
1.
In ion exchange chromatography, proteins are separated based on
a.
Size
b.
Size and shape
c.
Specific binding affinity
d.
Charge
2.
In gel filtration chromatography, proteins are separated based on
a.
S Size
b.
Size and shape
c.
Specific binding affinity
d.
Charge
3.
What type of chromatography can you use to determine quaternary structure of a protein?
a.
Ion exchange
b.
Gel filtration
c.
Affinity
4.
In gel filtration chromatography
a.
Largest proteins elute last
b.
Smallest proteins elute last
c.
Positively charged proteins elute first
d.
Negatively charged proteins elute first
e.
Uncharged proteins elute last
5.
In anion exchange chromatography
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