MOLECULAR DIAGNOSTICS FINAL
EXAM QUESTIONS AND ANSWERS
100% ACCURATE
The advantages of using real-time PCR to detect Chlyamidia and Gonorrhea is... -
ANSWER-- allow detection of organisms that are hard to culture
- can be automated for high volume processing
MALDI-TOF improves traditional culture methods by... - ANSWER--reduces need for
biochem tests
-pathogen-same method for all bacteria and fungi
-cheaper cost per sample
Describe the precautions that the State lab takes to prevent contamination (3) -
ANSWER--designated clean (pre) and dirty (post) lab
-2 different hoods for set up and adding DNA before PCR
-hoods cleaned using UV light
Why not molecular methods? - ANSWER--cost
-requires technical skill--> resistant lab staff
-FDA approval
-standardized tests are not available
Hybrid Capture is used to detect... - ANSWER-HPV
How does Hybrid capture work? - ANSWER-- viral and human dna denatured (single
stranded)
- rna probe binds to viral dna
- antibody detects only double stranded dna
- single stranded dna is washed away
- second antibody with florescent label attaches to sandwich dna
- fluorescence is detected
Where on the growing nucleotide chain does the next nucleotide add? - ANSWER-
Carbon 3 of the ribose sugar
What enzyme adds the next nucleotide? - ANSWER-DNA polymase
What type of bond forms between the 2 nucleotides? - ANSWER-phosphodiester
Whats the complimentary sequence?
5' ATTCACGG 3' - ANSWER-3' AAGTGAAT 5'
, How does it change the set up for real time PCR? - ANSWER-molecular beacon
If you wanted to screen a new borne baby for multiple genetic disorders what test would
you use? - ANSWER-microarray
The 3 major components of that make up DNA? - ANSWER--sugar (ribose)
-phosphate group
-nitrogenous base
What is the overall charge of DNA and on what part is it found? - ANSWER-negative
phosphorus back bone
Name a method that uses the fact that DNA is negatively charged to work? -
ANSWER--PFGE
-agarose gel electrophoresis
What type of bonds are formed during hybridization when a strand of DNA finds its
compliment - ANSWER-Hydrogen?
What is an example of STR in DNA sequence? - ANSWER-AUG AUG AUG
2 difference of prok and euk transcription - ANSWER-1. monocistronic (euk) vs
polycistronic (prok) (rna can code for more than 1 protein)
2. prok has only 1 promotor, euk have many
What are the components of the t-rna structure? - ANSWER-- anticodon (bottom)
- amino acid (top)
What is unique about eukaryotic mRNA that allows it to be isolated from total RNA
extraction? - ANSWER-has a poly A tail to hybridize to
Define wobble position and how it relates to screening for SNPs in the human genome -
ANSWER-The last nucleotide can be interchanged but still produce the same amino
acid. That way a mistake can happen without affecting the product
What method is used to separate proteins by size? - ANSWER-SDS page
What are the major steps of DNA extraction? - ANSWER-1. cell lysis
a. physical - freeze then thaw, blending, beading
b. chemical - SDS page, proteinase K
2. separation - collum or phenol chloroform
3. concentration - add ethanol
What is the purpose of SDS page in DNA extraction? - ANSWER-detergent - breaks
down cell membarne
EXAM QUESTIONS AND ANSWERS
100% ACCURATE
The advantages of using real-time PCR to detect Chlyamidia and Gonorrhea is... -
ANSWER-- allow detection of organisms that are hard to culture
- can be automated for high volume processing
MALDI-TOF improves traditional culture methods by... - ANSWER--reduces need for
biochem tests
-pathogen-same method for all bacteria and fungi
-cheaper cost per sample
Describe the precautions that the State lab takes to prevent contamination (3) -
ANSWER--designated clean (pre) and dirty (post) lab
-2 different hoods for set up and adding DNA before PCR
-hoods cleaned using UV light
Why not molecular methods? - ANSWER--cost
-requires technical skill--> resistant lab staff
-FDA approval
-standardized tests are not available
Hybrid Capture is used to detect... - ANSWER-HPV
How does Hybrid capture work? - ANSWER-- viral and human dna denatured (single
stranded)
- rna probe binds to viral dna
- antibody detects only double stranded dna
- single stranded dna is washed away
- second antibody with florescent label attaches to sandwich dna
- fluorescence is detected
Where on the growing nucleotide chain does the next nucleotide add? - ANSWER-
Carbon 3 of the ribose sugar
What enzyme adds the next nucleotide? - ANSWER-DNA polymase
What type of bond forms between the 2 nucleotides? - ANSWER-phosphodiester
Whats the complimentary sequence?
5' ATTCACGG 3' - ANSWER-3' AAGTGAAT 5'
, How does it change the set up for real time PCR? - ANSWER-molecular beacon
If you wanted to screen a new borne baby for multiple genetic disorders what test would
you use? - ANSWER-microarray
The 3 major components of that make up DNA? - ANSWER--sugar (ribose)
-phosphate group
-nitrogenous base
What is the overall charge of DNA and on what part is it found? - ANSWER-negative
phosphorus back bone
Name a method that uses the fact that DNA is negatively charged to work? -
ANSWER--PFGE
-agarose gel electrophoresis
What type of bonds are formed during hybridization when a strand of DNA finds its
compliment - ANSWER-Hydrogen?
What is an example of STR in DNA sequence? - ANSWER-AUG AUG AUG
2 difference of prok and euk transcription - ANSWER-1. monocistronic (euk) vs
polycistronic (prok) (rna can code for more than 1 protein)
2. prok has only 1 promotor, euk have many
What are the components of the t-rna structure? - ANSWER-- anticodon (bottom)
- amino acid (top)
What is unique about eukaryotic mRNA that allows it to be isolated from total RNA
extraction? - ANSWER-has a poly A tail to hybridize to
Define wobble position and how it relates to screening for SNPs in the human genome -
ANSWER-The last nucleotide can be interchanged but still produce the same amino
acid. That way a mistake can happen without affecting the product
What method is used to separate proteins by size? - ANSWER-SDS page
What are the major steps of DNA extraction? - ANSWER-1. cell lysis
a. physical - freeze then thaw, blending, beading
b. chemical - SDS page, proteinase K
2. separation - collum or phenol chloroform
3. concentration - add ethanol
What is the purpose of SDS page in DNA extraction? - ANSWER-detergent - breaks
down cell membarne
