BIOL 2288 FINAL EXAM QUESTIONS WITH ANSWERS
100% CORRECT
ELISA relies on ___ to detect the presence of ___ in liquid samples - ANSWER antibodies
antigens
antibodies - ANSWER Specialized proteins that aid in destroying infectious agents
antigens - ANSWER Foreign material that invades the body
ELISA can detect minute amounts of what? (3) - ANSWER •Hormones (like hCG for
pregnancy tests)
•Drugs
•Disease agents
Steps of an ELISA - ANSWER 1.Add sample to wells and allow proteins (including
antigens) to bind (adsorb) to the plastic via hydrophobic interaction.
2.Add primary antibodies and allow to bind to bound antigen.
3.Add enzyme-labeled secondary antibodies and allow to bind to bound primary
antibodies.
4.Detect the presence of enzyme-labeled secondary antibodies by adding substrate.
Which of the following serves as the negative control in ELISA?
A.Sample from a potentially infected UTD student
B.Heat-killed disease agent suspended in phosphate buffered saline (PBS)
C.Phosphate buffered saline - ANSWER C.Phosphate buffered saline
ELISA - Why do we do this in duplicate? - ANSWER To make sure there isn't conflicting
,data. It provides confidence, as we can reproduce results. Makes sure technique is
correct as well.
Can also do triplicate
Proteins Bind Plate By ___Interaction - ANSWER Hydrophobic
Antigen (if present) Binds to what? - ANSWER Plate
T or F: The antigen of interest will be present in the serum sample from an uninfected
student - ANSWER False
Wash procedure is done ___ the first two times with ___ - ANSWER twice with PBS buffer
and tween20 (a detergent)
Primary Antibody Binds to what?
(if present) - ANSWER Antigen
In an ELISA performed on a sample from an uninfected student, the primary antibody
will bind to
A.The plastic
B.The antigen
C.The phosphate buffered saline
D.None of the above - ANSWER D.None of the above
The ___ is conjugated with an enzyme - ANSWER secondary antibody
Secondary Antibody Binds to what?
(if present) - ANSWER Primary Antibody
, The wash procedure after adding secondary antibody is done ___ times - ANSWER three
Always tap out water before adding more - ANSWER
Substrate Interacts With ___ on Secondary (if present) - ANSWER Enzyme
The enzyme takes TMB, which is colorless and convert TMB into a colored product if
present. What color in this ELISA experiment? - ANSWER Blue
Primers are about how many nucleotides in length? - ANSWER 20
Primers Must be complementary to the ___ end of the gene to be amplified, specifically
for the gene - ANSWER 3'
Primers cannot bind to bacterial DNA because they code for specific genes, only to the
human to extend it in the 5' - > 3' direction (region to be amplified read in 3- ->5'
direction) - ANSWER
Excess quantity of primer in PCR rxn, why? - ANSWER Bc lots of DNA will be
synthesized, need to have enough primers.
What is the optimal temperature of Taq DNA polymerase? - ANSWER 72 - 75 degrees C
Human nuclear DNA vs. Human mtDNA - ANSWER Nuclear - very large, very complex,
very long linear chains, two copies of a given nuclear gene per cell (one maternal and
one paternal)
mtDNA -very small, comparatively simple, small circular loop, many copies per cell
(each cell has many mitochondria which has many copies of mtDNA and are maternal)
What is the expected size of this PCR product (Left primer:15971, Right primer:16411)
and total: 16,569 bp? - ANSWER 440
100% CORRECT
ELISA relies on ___ to detect the presence of ___ in liquid samples - ANSWER antibodies
antigens
antibodies - ANSWER Specialized proteins that aid in destroying infectious agents
antigens - ANSWER Foreign material that invades the body
ELISA can detect minute amounts of what? (3) - ANSWER •Hormones (like hCG for
pregnancy tests)
•Drugs
•Disease agents
Steps of an ELISA - ANSWER 1.Add sample to wells and allow proteins (including
antigens) to bind (adsorb) to the plastic via hydrophobic interaction.
2.Add primary antibodies and allow to bind to bound antigen.
3.Add enzyme-labeled secondary antibodies and allow to bind to bound primary
antibodies.
4.Detect the presence of enzyme-labeled secondary antibodies by adding substrate.
Which of the following serves as the negative control in ELISA?
A.Sample from a potentially infected UTD student
B.Heat-killed disease agent suspended in phosphate buffered saline (PBS)
C.Phosphate buffered saline - ANSWER C.Phosphate buffered saline
ELISA - Why do we do this in duplicate? - ANSWER To make sure there isn't conflicting
,data. It provides confidence, as we can reproduce results. Makes sure technique is
correct as well.
Can also do triplicate
Proteins Bind Plate By ___Interaction - ANSWER Hydrophobic
Antigen (if present) Binds to what? - ANSWER Plate
T or F: The antigen of interest will be present in the serum sample from an uninfected
student - ANSWER False
Wash procedure is done ___ the first two times with ___ - ANSWER twice with PBS buffer
and tween20 (a detergent)
Primary Antibody Binds to what?
(if present) - ANSWER Antigen
In an ELISA performed on a sample from an uninfected student, the primary antibody
will bind to
A.The plastic
B.The antigen
C.The phosphate buffered saline
D.None of the above - ANSWER D.None of the above
The ___ is conjugated with an enzyme - ANSWER secondary antibody
Secondary Antibody Binds to what?
(if present) - ANSWER Primary Antibody
, The wash procedure after adding secondary antibody is done ___ times - ANSWER three
Always tap out water before adding more - ANSWER
Substrate Interacts With ___ on Secondary (if present) - ANSWER Enzyme
The enzyme takes TMB, which is colorless and convert TMB into a colored product if
present. What color in this ELISA experiment? - ANSWER Blue
Primers are about how many nucleotides in length? - ANSWER 20
Primers Must be complementary to the ___ end of the gene to be amplified, specifically
for the gene - ANSWER 3'
Primers cannot bind to bacterial DNA because they code for specific genes, only to the
human to extend it in the 5' - > 3' direction (region to be amplified read in 3- ->5'
direction) - ANSWER
Excess quantity of primer in PCR rxn, why? - ANSWER Bc lots of DNA will be
synthesized, need to have enough primers.
What is the optimal temperature of Taq DNA polymerase? - ANSWER 72 - 75 degrees C
Human nuclear DNA vs. Human mtDNA - ANSWER Nuclear - very large, very complex,
very long linear chains, two copies of a given nuclear gene per cell (one maternal and
one paternal)
mtDNA -very small, comparatively simple, small circular loop, many copies per cell
(each cell has many mitochondria which has many copies of mtDNA and are maternal)
What is the expected size of this PCR product (Left primer:15971, Right primer:16411)
and total: 16,569 bp? - ANSWER 440
