Please answer the following multiple-part question regarding vectored cloning:
a) A cloning vector must _________.
A. be a circular dsDNA molecule
B. contain a replication origin
C. contain an multicloning site (MCS)
D. contain marker genes
b) Vectored cloning is superior to PCR cloning of a gene in terms of __________.
A. time it takes to finish cloning
B. size of DNA that can be cloned
C. amounts of cloned DNA that can be generated
D. ease of isolation of the DNA fragment to be cloned
c) Which of the following is a recombinant DNA molecule?
A. pUC DNA digested by EcoRI
B. Fish insulin gene, PCR amplified and then cut by EcoRI
C. pUC18 DNA and fish insulin DNA ligated by a DNA liagse
D. pUC DNA and fish insulin gene DNA, both cut by EcoRI and mixed
d) You may insert a foreign DNA fragment inside a live cell using each of the following methods except________.
A. inject the DNA inside the cell
B. mix the DNA with the cell and freeze the mixture in liquid nitrogen
C. mix DNA with the cell and momentarily heat the cell then cool down
D. mix DNA with the cell and momentarily zap the cells with a high voltage
E. cover the DNA in a lipid micelle and mix the micelle with the cell
e) Technically, the ampicillin resistance gene is a selectable marker gene but the beta lacZ gene or the green fluorescent protein (GFP) gene is a reporter gene of a cloning vector. What is the difference between a selectable marker and a reporter gene?
A. A selectable marker is a bacterial gene, a reporter gene is a eukaryotic gene
B. Marker genes are used in bacterial system; reporter genes are used in eukaryotic system.
C. The selectable markers help eliminate cells lacking the maker, reporter genes do not offer this option.
Please answer the following multiple-part question regarding vectored cloning:
a) A cloning vector must _________.
A. be a circular dsDNA molecule
B. contain a replication origin
C. contain an multicloning site (MCS)
D. contain marker genes
b) Vectored cloning is superior to PCR cloning of a gene in terms of __________.
A. time it takes to finish cloning
B. size of DNA that can be cloned
C. amounts of cloned DNA that can be generated
D. ease of isolation of the DNA fragment to be cloned
c) Which of the following is a recombinant DNA molecule?
A. pUC DNA digested by EcoRI
B. Fish insulin gene, PCR amplified and then cut by EcoRI
C. pUC18 DNA and fish insulin DNA ligated by a DNA liagse
D. pUC DNA and fish insulin gene DNA, both cut by EcoRI and mixed
d) You may insert a foreign DNA fragment inside a live cell using each of the following methods except________.
A. inject the DNA inside the cell
B. mix the DNA with the cell and freeze the mixture in liquid nitrogen
C. mix DNA with the cell and momentarily heat the cell then cool down
D. mix DNA with the cell and momentarily zap the cells with a high voltage
E. cover the DNA in a lipid micelle and mix the micelle with the cell
e) Technically, the ampicillin resistance gene is a selectable marker gene but the beta lacZ gene or the green fluorescent protein (GFP) gene is a reporter gene of a cloning vector. What is the difference between a selectable marker and a reporter gene?
A. A selectable marker is a bacterial gene, a reporter gene is a eukaryotic gene
B. Marker genes are used in bacterial system; reporter genes are used in eukaryotic system.
C. The selectable markers help eliminate cells lacking the maker, reporter genes do not offer this option.
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Why does a template-dependent DNA polymerase require a primer to initiate DNA synthesis?
DNA polymerase require 5' phosphate to add a new nucleotide | |
DNA polymerase require 3' hydroxyl group to add a new nucleotide |
DNA polymerase require energy by hydrolizing the primers |
Which of the following enzymes is template independent?
Reverse transcriptase | |
Klenow fragment |
DNA polymerase | |
Terminal deoxynucleotidyl transferase |
RNA polymerase |
The natural function of a DNA ligase enzyme is:
Joining together DNA sequences with compatible cohesive ends from two different organisms to yield a recombinant DNA molecule | |
Sealing single stranded nicks in double stranded DNA molecules |
Joining together two blunt ended fragments of DNA within a cell | |
Joining together two single stranded DNA molecules |
The natural function of 3' -> 5' exonuclease activity of a DNA polymerase is to:
Remove the 5' end of the DNA strand that is being copied | |
Remove damaged nucleotide from the template strand |
Remove nucleotides from the end of DNA to generate blunt ends | |
Remove incorrect nucleotides from the newly synthesized DNA |
Which technique is used to resolve the different sizes of DNA fragments?
Gene cloning | |
PCR |
DNA sequencing | |
Gel electrophoresis |
Which of the following vectors does not contain bacterial origin of replication (oriC)?
plasmid | |
cosmid |
bacterial artificial chromosome | |
lambda vector |
Which process of bacteriophage is not used for gene cloning?
Concatemer formation | |
Lytic cycle |
Lysogenic cycle | |
viral packaging |
What vector would be best suited for creating a contig of bovine (cattle) chromosome 10?
lambda vector | |
Cosmid vector |
P1 vector | |
Plasmid |
E. coli takes up plasmid DNA by which of the following methods?
Transduction | |
Transformation |
Conjugation | |
Transfection |
The following times and temperatures are an example of the steps for PCR. You can use the Figure to help you answer the following questions.
Why is the first step is carried out at 94°C?
To degrade the template | |
To denature the template |
To activate the polymerase | |
To facilitate the primer binding |
What happens in the reaction when the temperature shifts to 55°C during cycling?
the DNA polymerase will carry out DNA synthesis by extending the annealed primers | |
the DNA polymerase will finish DNA synthesis |
the primers anneal to the single-stranded regions of the DNA | |
the primers anneal to the double-stranded regions of the DNA |
During cycling, what occurs when the temperature is at 72°C?
the primers anneal to the double-stranded regions of the DNA | |
the DNA polymerase will carry out DNA synthesis by extending the annealed primers |
the primers anneal to the single-stranded regions of the DNA | |
the DNA polymerase will finish DNA synthesis |
You designed a set of primers to amplify 1 kb DNA with polymerase chain reaction. By which cycle of polymerase chain reaction, we would expect to see the first double stranded DNA with expected size?
The end of cycle 1 | |
The end of cycle 2 |
The end of cycle3 | |
The end of cycle 4 |
Which of the following did Erwin Chargaff observe?
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The melting temperature (Tm) of a DNA duplex is:
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Given your knowledge and the descriptions of each of thesemethods, which of the following does NOT rely on the ability ofnucleic acids to hybridize with each other?
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Which of the following is a possible reason why DNA uses thymineinstead of uracil?
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Which of the following is an enzyme used in cloning to breakcovalent bonds?
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Which of the following could you use to remove the 5' phosphatesafter cleavage of a plasmid with a single type of restrictionenzyme to ensure that the plasmid would not simply fuse backtogether upon addition of ligase?
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Which of the following would you use to create a cDNA librarythat you would not use for a genomic library?
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A plasmid vector typically has which of the followingfeatures?
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Which of the following is FALSE of genomic or cDNAlibraries?
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Which of the following is NOT a typical component of apolymerase chain reaction (PCR)?
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How can a fusion protein be formed?
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Why is green fluorescent protein (GFP) so useful in visualizingfusion proteins in eukaryotes?
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You wish to determine the cellular location of a protein butunfortunately it is not particularly antigenic (can t develop astrong antibody response). How could you solve this problem?
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A positive result for the yeast two-hybrid analysis means thefollowing:
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Which of the following is the approximate size of the humangenome?
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Which of the following is correct about the structure orlocation of genes?
Question 22 options:
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