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BCH5413 Exam 1 Questions With Complete Solutions
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BCH5413 Exam 1 Questions With Complete Solutions
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BCH5413 Exam 1 Questions With Complete Solutions1. Choosing an E. coli protein expression system has a lot of
advantages such as yield, cost and ease.
Discuss (not just list) two reasons why a researcher might not
choose this type of system.
Be specific, what might their goal be and why wouldn't this
system work for them? Correct Answer 1. no post-translational
modification - E. coli system is not sophisticated enough for
downstream experimental goals
2. limited cDNA size / codon usage differences - limit the vector
choice for preferred protein
--- "experimental difficulty" = protein expression may not
always be the same between E. coli + mammals
1. In a PCR reaction, why do you need:
a. Both forward + reverse primer
,b. Template
c. dNTP
d. Magnesium
e. DNA polymerase Correct Answer a. provide a "free" 3'-OH
group to which the DNA
polymerase can add dNTPs
b. to make copies of
c. building block nucleotides
d. co-factor = increase DNA polymerase in replicating the
template DNA
e. add nucleotides to new strand
1. In cDNA library construction:
a. What is cDNA?
b. why would an investigator want to create a cDNA library?
,c. what is the advantage of using beads containing oligo T
nucleotides for isolating the RNA? Correct Answer a.
complementary DNA - made via reverse transcriptase
b. to obtain a set of clones representing all mRNAs expressed in
a given cell type
c. oligo T nucleotides = primer to bind to the 3' end poly(A) tail
of most eukaryotic mRNAs
1. In gel electrophoresis:
a. why does DNA move toward the positive pole?
b. why does DNA move according to size? Correct Answer a.
DNA = negative charge move towards positive charge
b. smaller fragments move faster through matrix towards
positive charge
1. In Maxam & Gilbert Sequencing: (chemical modification +
cleavage)
, a. Why do you need to radioactively label the fragment that is
being sequenced?
b. Why do you need to chemically modify the bases?
c. Why do you need an polyacrylamide gel?
d. Why is this method considered direct as compared to other
methods? Correct Answer a. To read the sequence on the gel
b. So that the DNA can be cleaved at SPECIFIC bases
c. Allows DNA to be separated by size by a SINGLE nucleotide
d. the DNA to be sequenced is broken up into readable
fragments
1. In the experiment described that supports DNA as the genetic
material:
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