BIOL 2070- RESEARCH METHODS FINAL EXAM QUESTIONS
AND ANSWERS WITH COMPLETE SOLUTIONS VERIFIED
You have a solution of protein that has a concentration of 200.2 ng/mL. What is
the concentration in micrograms/L?
200.2
You want to make a 125mL solution with a final concentration of 250mM. The
stock solution is 0.5M how much stock solution is needed?
0.0625L
Primary piece of literature. What are some ways to identify?
Firsthand report of an experiment that was conducted by the authors
- Speaking from experience
- Methods/materials, results section
- direct report of experiment conducted
Tertiary piece of literature
A mix of primary and secondary literature
How long should a primer be?
15-18 nucleotides long - complimentary sequence
How do you design a forward primer with a restriction endonuclease sites?
,Add some random nucleotides preceding the site, add the site at the 5' end of the
primer and then add the sequences that are complementary to the DNA being
sequenced
How do you design a reverse primer?
The reverse complement of the DNA sequence provided
A yeast-1-hybrid system that cannot grow on histidine-deficient media. The bait
and prey bind to each other. What experimental result is consistent with this
claim?
Growth on histidine-deficient media
A yeast-1-hybrid system that cannot grow on histidine-deficient media. The bait
and prey do not bind to each other. But there was a mutation in the yeast RNA
polymerase and it's able to bind the activation domain. What experimental result
is consistent with this claim?
No growth on histidine-deficient media
Yeast-2-hybrid system that cannot grow on histidine-deficient media. Bait and
prey bind to each other but the yeast RNA polymerase can also bind to the
activation domain. What experimental results are consistent with the
information?
Growth on histidine-deficient media
False positive result
The RNA polymerase binds to the DNA directly so it transcribes but the bait and prey do
not bind
False negative result
, DNA binding domain prevents it from being imported into the nucleus but the bait and
prey bind
What is a construct?
Artificially engineered segments of DNA that are created by combining different DNA
sequences
How would you purify a protein using a Nickel agarose column?
By fusing a his tag to the protein product - flood the column with thrombin to elute
protein of interest (disrupts the binding between his tag and nickel)
Which restriction sites should be used to clone a gene if you want to fuse the his
tag?
Use restriction sites DOWNSTREAM of the his tag - ensures his tag is not removed
from vector
What is important about the BL21 strain of competent cells?
A chemically engineered E. Coli strain that is deficient in cytoplasmic protease and
membrane protease which allow transformation and expression of genes from another
cell-type
How to read the sequence from a diagram?
The sequence provided in the 5'-3' direction is the reverse complement of the original
sequence
What is the purpose of agarose gel electrophoresis?
To determine the size of DNA fragments. Smaller fragments move through the gel faster
Would a kidney cell and a brain cell from the same person have all the same
genes?
AND ANSWERS WITH COMPLETE SOLUTIONS VERIFIED
You have a solution of protein that has a concentration of 200.2 ng/mL. What is
the concentration in micrograms/L?
200.2
You want to make a 125mL solution with a final concentration of 250mM. The
stock solution is 0.5M how much stock solution is needed?
0.0625L
Primary piece of literature. What are some ways to identify?
Firsthand report of an experiment that was conducted by the authors
- Speaking from experience
- Methods/materials, results section
- direct report of experiment conducted
Tertiary piece of literature
A mix of primary and secondary literature
How long should a primer be?
15-18 nucleotides long - complimentary sequence
How do you design a forward primer with a restriction endonuclease sites?
,Add some random nucleotides preceding the site, add the site at the 5' end of the
primer and then add the sequences that are complementary to the DNA being
sequenced
How do you design a reverse primer?
The reverse complement of the DNA sequence provided
A yeast-1-hybrid system that cannot grow on histidine-deficient media. The bait
and prey bind to each other. What experimental result is consistent with this
claim?
Growth on histidine-deficient media
A yeast-1-hybrid system that cannot grow on histidine-deficient media. The bait
and prey do not bind to each other. But there was a mutation in the yeast RNA
polymerase and it's able to bind the activation domain. What experimental result
is consistent with this claim?
No growth on histidine-deficient media
Yeast-2-hybrid system that cannot grow on histidine-deficient media. Bait and
prey bind to each other but the yeast RNA polymerase can also bind to the
activation domain. What experimental results are consistent with the
information?
Growth on histidine-deficient media
False positive result
The RNA polymerase binds to the DNA directly so it transcribes but the bait and prey do
not bind
False negative result
, DNA binding domain prevents it from being imported into the nucleus but the bait and
prey bind
What is a construct?
Artificially engineered segments of DNA that are created by combining different DNA
sequences
How would you purify a protein using a Nickel agarose column?
By fusing a his tag to the protein product - flood the column with thrombin to elute
protein of interest (disrupts the binding between his tag and nickel)
Which restriction sites should be used to clone a gene if you want to fuse the his
tag?
Use restriction sites DOWNSTREAM of the his tag - ensures his tag is not removed
from vector
What is important about the BL21 strain of competent cells?
A chemically engineered E. Coli strain that is deficient in cytoplasmic protease and
membrane protease which allow transformation and expression of genes from another
cell-type
How to read the sequence from a diagram?
The sequence provided in the 5'-3' direction is the reverse complement of the original
sequence
What is the purpose of agarose gel electrophoresis?
To determine the size of DNA fragments. Smaller fragments move through the gel faster
Would a kidney cell and a brain cell from the same person have all the same
genes?
