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Waksman Project Unit Test 1

Multiple Choice
Identify the choice that best completes the statement or answers the question.
 
 
nar001-1.jpg
 

 1. 
The process illustrated in the above diagram is ----
a.
transformation- bacterial cell are induced to uptake a new plasmid.
d.
asexual reproduction- bacterial cells are replicating and dividing
b.
transillumination- bacterial cells are made competent.
e.
sexual reproduction- bacterial cells are exchanging genes.
c.
horizontal transfer of genetic material between bacterial cells.
 
 
The following questions refer the series of diagrams below. Numbers 1, 3, and 5 refer to the structures in the diagram. Number 2, 4, and 6 refer to step in the process of restriction digest of a plasmid
nar002-1.jpg
 

 2. 
How many restriction enzymes are required for the digest of the plasmid reaction and uptake of the insert to be sucessful? How many restriction sites are necessary?
a.
1 restriction enzyme......1 restriction site
b.
2 different restriction enzymes......1 restriction site- This could work if a selectable marker gene is present on the plasmid
c.
3 different restriction enzymes.....1 restriction site- this could work but only if the fragments created were of the same size.
d.
2 different restriction enzymes.....2 different restriction sites- but the sites must be close together.
e.
both A and D could have worked with this example
 

 3. 
The step between 3 and 4 on the diagram shows an opened plasmid. Which statement best describes how the plasmid was opened?
a.
restriction digest with restriction enzymes
b.
PCR- polymerase chain reaction
c.
mini prep- to remove the plasmid
d.
reverse transcriptase
e.
none of the above
 
 
The following gel show a lane of marker ladder then one plasmid with uncut and cut lanes and a second, different plasmid also with an uncut and cut lane.  The extra band in the uncut lanes are the plasmid in slightly different forms- supercoiled and nicked.
nar003-1.jpg
 

 4. 
Which are logical conclusions from the first set of Uncut and Cut lanes based on the observable bands in the gel?
a.
There is no insert. The smallest fragment in the cut lane is extra space where the PCR primers amplify the backbone.
b.
There is a large insert approximately 4000 bases in size.  The smallest fragment in the cut lane is primer dimer
c.
There is small insert of less then 100 base not visible on the gel. The small visible fragment in the cut lane is the primer dimer
d.
There is contamination and no evidence of a successful digestion is visible.
e.
There is an internal restriction site on the insert.
 

 5. 
Compare both CUT lanes on the gel. The explanation for the large bands at 4,000 bases is best described by which of the following?
a.
both bands are approximately 2,000 bases long.
b.
Both bands are are uncut, whole plasmid
c.
Both bands are evidence of contamination
d.
Both bands show that the “inserts” are of the same size
e.
Both bands are vector backbone from the original plasmid
 
 
nar004-1.jpg
 

 6. 
Observe clone number 1.  Which observation(s) can be made about the gel electrophoresis of this clone?
a.
The clone has an insert of approximately 300 base pairs.(+/- 50  bases)
b.
The clone shows contamination- most likely from picking two colonies from the plate that were two close together.
c.
The clone shows a PCR product of approximately 500 bases.
d.
 The digest lane and the PCR lane “agree” about the size of the insert.
e.
Both a and d are correct
 

 7. 
Consider clone number 4.  Compare the cut and the PCR lanes. What are possible observations regarding this clone?
a.
There is single large insert with and an internal restriction site on the insert.
b.
There is a single large insert
c.
There is contamination- the PCR lane confirms this with two different sized fragments
d.
There is a single large insert and primer dimer in the cut lanes
e.
bothe a and d are correct
 
 
nar005-1.jpg
 

 8. 
The above image shows a plasmid (with no insert) digest with Ava I enzyme on three different plasmids.
Which of the following explanation best describes the results observed from the gel.
a.
Lane 1 shows a plasmid with possibly one cut site
b.
Lane 1 shows a plasmid with two possible cut sites
c.
Lane 2 shows a digest with two cut sites
d.
Lane 3 shows a plasmid with three cut sites
e.
All of the above are correct
 
 
Pipettor Practice:

nar006-1.jpg
 

 9. 
Which of the following should be used for pipetting 183.4ul?
a.
1 only
b.
2 and 3
c.
3 only
d.
3 and 4
e.
4 only
 
 
Pipettor Window:

nar007-1.jpg

nar007-2.jpg
 

 10. 
Which of pipettor would be ideal for pipette volume #6
a.
1
b.
2
c.
3
d.
4
e.
none of the above
 

 11. 
Which pipettor would be ideal to pipette volume #5?
a.
1
b.
2
c.
3
d.
4
e.
none of the above
 
 
2nar008-1.jpg
 

 12. 
Which lane has the biggest insert?
a.
1
d.
6
b.
2
e.
7
c.
4
 

 13. 
Which lane shows the smallest insert?
a.
1
d.
6
b.
2
e.
7
c.
3
 

 14. 
Lane five can best described as a ---
a.
failed PCR reaction
b.
a failed restriction digest
c.
a poor mini prep
d.
both a and c
e.
all a, b, and c
 

 15. 
Which of the following could be an incorrect statement?
a.
Lane 5 possibly forgot to add primers
b.
Lane1 had a succesful PCR and alot of template DNA in the reaction.
c.
Lane 2’s insert is about 500 bases long
d.
The fragment in lane 7 is about 2000 bp long
e.
The results of lane 4 show an insert of about 200 bases
 
 
nar009-1.jpg
 

 16. 
Which lanes show a restriction digest?
a.
Lanes 1, 2, 3 and 4 all show the products of a restriction digest.
b.
Lane 1, 3, 5 al show the products of a restriction digest
c.
Lane 2, 4, and 6 all show the products of a restriction digest
d.
All the lanes show a restriction digest
e.
None of the lane show a restriction digest
 

 17. 
Which the correct explanation for the bands in the lane labeled 1?
a.
Contamination of mutiple plasmids
b.
Whoever made the reaction forgot to add primers
c.
There is an unut plasmid but it is running in three different forms.
d.
There is a restriction digest with several fragments due to the digest
e.
No DNA
 

 18. 
What is the best explanation for lane 4?
a.
Uncut plasmid running in multiple forms
b.
Internal restriction site in the cDNA
c.
too much DNA that is smearing in the lane
d.
forgot to add primers
e.
all of the above are possible
 
 
nar010-1.jpg
 

 19. 
Assuming a successful transformation of each plasmid into a bacterial cell. Which will allow the bacterial cell to grow on a plate of LB only?
a.
the cell with plasmid 1 will not grow since it does not have insert
b.
the cell with plasmid 2 will grow since there is a cDNA present
c.
Both types of cell (one with plasmid 1 and the other with plasmid 2) will grow since there is no selection pressure on the plate
d.
All cells, regardless of whether they picked up the plasmid will grow since there is no amp in the plate.
e.
Both C and D are correct
 

 20. 
When using gel electrophoresis, which of the following is FALSE?
a.
Fragments will separate by size; the largest fragments will move the least
b.
the sample will migrate through an agarose gel
c.
the higher the voltage the faster the sample will move through the gel
d.
DNA is a negatively charged particle so it will need to be away from the RED or + end
e.
All of the above are true
f.
All of the above are false
 



 
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