Module 2 stuff Flashcards
All of the following are potentially good sources of genomic DNA EXCEPT:
a. solid tissue
b. WBCs
c. mature RBCs
d. buccal cells
mature RBCs
After isolating the DNA specimen, you assess the quality of the specimen prior to amplification. The mass spectrometry results read at 280 nm. This result suggests that:
the specimen is contaminated with left-over proteins
What is the key enzyme used in a polymerase chain reaction (PCR)?
polymerase
In real-time PCR:
a. the amount of signal generated is directly proportional to the amount of template
b. RNA targets are used in an isothermal reaction.
c. an isolated mRNA and Thermus thermophilus is used.
d. RNA:DNA hybrids are bound to a solid phase.
the amount of signal generated is directly proportional to the amount of template
The annealing step in a PCR involves:
a. adding nucleotides to primed sites of the DNA strands.
b. binding of the primers to the single-stranded DNA.
c. unwinding and separating the double-stranded DNA.
d. making a cDNA from an mRNA strand.
binding of the primers to the single-stranded DNA
During the extension phase of PCR:
a. oligonucleotide primers attach at ends of the strands to promote replication
b. the dsDNA separates
c. tRNA delivers a nucleotide to the appropriate position
d. heat-stable DNA polymerase synthesizes complementary strands
heat-stable DNA polymerase synthesizes complementary strands
False positive results in molecular testing are most likely caused by which one of the following?
a. Inhibitors present in the sample
b. Using too little DNA
c. Contamination with outside DNA
d. Excessive washing
Contamination with outside DNA
You want to determine the accuracy of hybridization of a probe to a Southern blot to eliminate binding of the probe to incorrect target sequences. Which one of the following would be an appropriate positive control to assess assay sensitivity?
a. Water
b. Sequences complementary to the probe
c. Sequences from a housekeeping gene such as beta actin
d. Sequences that are not complementary to the probe
Sequences complementary to the probe
The purpose of restriction fragment polymorphism analysis (RFLP), which is the methodology of the Southern Blot, is to:
identify the presence of mutations or sequence changes
You have prepared a restriction enzyme digest of DNA from an individual who might have a genetic disorder. On the gel, you note that the digest from this individual produces one fragment, whereas the digest from a normal healthy individual produces two fragments. All controls worked correctly and you used the same reagents for all testing. What is your interpretation?
The diseased individual’s DNA is missing a restriction site (a recognition sequence).
In a pyrosequencing reaction, a single strand of DNA is used as a template to synthesize the complimentary strand. The instrument uses ___ to determine the results.
chemiluminescence
The enzyme used to cleave nucleotides one at a time from the ends is:
an exonuclease
The restriction endonuclease EcoRI recognizes ___ and produces a ___.
GAATTC ; staggered cut
___ degrades nucleic acid.
nucleases
___ catalyzes the synthesis of DNA from RNA or DNA templates.
reverse transcriptase
___ covalently joins 2 DNA strands.
ligases
What is the method of amplification used for this type of methodology?
Reverse transcriptase PCR
target
What is the method of amplification used for this type of methodology?
Rolling circle amplification
probe
What is the method of amplification used for this type of methodology?
Transcription mediated amplification
target
What is the method of amplification used for this type of methodology?
Polymerase chain reaction
target
What is the method of amplification used for this type of methodology?
Branched DNA
signal
What is the method of amplification used for this type of methodology?
Serial invasive signal amplification
signal
What is the method of amplification used for this type of methodology ?
Ligase chain reaction
target
Name the enzyme(s) required for this type of methodology:
Reverse transcriptase PCR
- reverse transcriptase
2. DNA polymerase
Name the enzyme(s) required for this type of methodology:
Ligase chain reaction
DNA ligase
Name the enzyme(s) required for this type of methodology:
WGA
processive DNA polymerase
Name the enzyme(s) required for this type of methodology:
LAMP
DNA polymerase
Name the enzyme(s) required for this type of methodology:
PCR
DNA polymerase
A restriction enzyme recognizes the sequence:
5’CT ^ ATAG 3’
…and cuts as indicated by the “^”
What should be the complementary strand?
3’ GATA ^ TC 5’
After 15 cycles of PCR, what is the estimated product of dsDNA?
32,768
If a DNA probe is added to nitrocellulose after the transfer step but without performing the blocking step, which of the following will occur?
a. The probe will bind to the target.
b. Bound probe will be washed away in the next step.
c. The DNA target on the nitrocellulose will be unable to bind the probe.
d. Unoccupied spaces on the nitrocellulose will bind the probe.
Unoccupied spaces on the nitrocellulose will bind the probe.
In the presence of salt, DNA is precipitated out from solution by:
Alcohols, such as 95% ethanol or isopropanol
For the purpose of diagnosing a genetic disease, which of the following is the most useful component of whole blood to be used for the extraction of DNA?
a. Platelets
b. Plasma
c. Red blood cells
d. Leukocytes
Leukocytes
In RNA, which nucleotide base replaces thymine of DNA?
uracil
Which of the following regarding Loop-mediated amplification (LAMP) is NOT true?
a. LAMP uses a strand displacement methodology.
b. The LAMP procedure makes looped structures of differing sizes.
c. The LAMP procedure is isothermal.
d. LAMP’s main enzyme used is RNA ligase.
LAMP’s main enzyme used is RNA ligase
All of the following are molecular diagnostic techniques that do NOT require electrophoresis EXCEPT:
a. Matrix-Assisted Laser-Desorption Ionization Time-of-flight (MALDI-TOF)
b. HIgh-throughput sequencing
c. Southern Blot
d. Pyrosequencing
Southern Blot
What refers to a genetic element that does not code for a functional gene product?
pseudogenes
On a Southern Blot, what happens if the washing step is insufficient?
It may cause a high background and non-interpretable results
A blot which uses a DNA probe produced by reverse transcription of the messenger RNA is ___.
Northern Blot
What is the correct temperature range for this step of PCR?
Denaturalization
94–95 degrees C
What is the correct temperature range for this step of PCR?
Extension of DNA
72 degrees C
What is the correct temperature range for this step of PCR?
Primer annealing
55–65 degrees C
Thermus aquaticus, a microbe found in hot springs, is utilized in the lab to as ___.
Taq DNA polymerase
What does Serial Invasive Signal Amplification do?
generates fluorescence to amplify the signal
