Podcast
Questions and Answers
What enzyme is needed for the synthesis of a DNA strand?
What enzyme is needed for the synthesis of a DNA strand?
- Protease
- ACE enzyme
- Taq DNA polymerase (correct)
- Taq RNA polymerase
At what temperature does primer hybridization occur?
At what temperature does primer hybridization occur?
- 72 degree
- 70 degree
- 40-60 degree (correct)
- 96 degree
What genotype should be in people who have higher levels of the ACE enzyme in their blood.
What genotype should be in people who have higher levels of the ACE enzyme in their blood.
- ll
- Il and DD
- DD (correct)
- There are no correct answer
Indicate which reagent is missing from the PCR reaction mixture listed:
The PCR reaction mixture contains: PCR reaction buffer (consisting of KCl, Tris, and MgCl2), DNA Taq polymerase enzyme responsible for the synthesis of a new DNA strand from bacteria isolated from Thermus aquaticus, 25 mM MgCl2, DNA, and primer.
Indicate which reagent is missing from the PCR reaction mixture listed: The PCR reaction mixture contains: PCR reaction buffer (consisting of KCl, Tris, and MgCl2), DNA Taq polymerase enzyme responsible for the synthesis of a new DNA strand from bacteria isolated from Thermus aquaticus, 25 mM MgCl2, DNA, and primer.
A variant of the ACE gene allele with Alu insertion can be identified by which of the following methods?
A variant of the ACE gene allele with Alu insertion can be identified by which of the following methods?
What is the ratio of genotypes according to Hardy-Weinberg's law (ACE gene alleles)?
What is the ratio of genotypes according to Hardy-Weinberg's law (ACE gene alleles)?
Indicate which reagent is missing from the PCR reaction mixture listed:
The PCR reaction mixture contains: PCR reaction buffer (consisting of KCI, Tris and MgCI2), deoxyribonucleoside
triphosphates (dNTPs -ie a mixture of dATP, dCTP, dGTP and dITP), DNA Taq polymerase (an enzyme responsible for
the synthesis of a new DNA strand from bacteria isolated from (Thermus aquaticus), 25 mM MgCI2 and DNA tested.
Indicate which reagent is missing from the PCR reaction mixture listed: The PCR reaction mixture contains: PCR reaction buffer (consisting of KCI, Tris and MgCI2), deoxyribonucleoside triphosphates (dNTPs -ie a mixture of dATP, dCTP, dGTP and dITP), DNA Taq polymerase (an enzyme responsible for the synthesis of a new DNA strand from bacteria isolated from (Thermus aquaticus), 25 mM MgCI2 and DNA tested.
List the stages of PCR reaction.
List the stages of PCR reaction.
Specify the size of the Alu sequence
Specify the size of the Alu sequence
During DNA denaturation
During DNA denaturation
evaluate the pcr reaction resulat according to the figure provided and identify the genotype
evaluate the pcr reaction resulat according to the figure provided and identify the genotype
at what concentrntrantion in the gel are ampliged DNA fragment
at what concentrntrantion in the gel are ampliged DNA fragment
Which of the following is not required in order to carry out PCR reaction? Select one
Which of the following is not required in order to carry out PCR reaction? Select one
How long does for electrophoresis take to detect the 1 and D aileles of
the ACE gene?
How long does for electrophoresis take to detect the 1 and D aileles of the ACE gene?
what is the size of the fragment amplified by ACE gene allele with a deletion
what is the size of the fragment amplified by ACE gene allele with a deletion
DNA fragments that are S 00 bp, 000
S
and 000 bp
length are separated by gell electrophoresis Which
fragment will migrate farthest in the get?
DNA fragments that are S 00 bp, 000 S and 000 bp length are separated by gell electrophoresis Which fragment will migrate farthest in the get?
Which type of reagent do not need to be added into PCR reaction? Select one:
Which type of reagent do not need to be added into PCR reaction? Select one:
hat componemts s hould be added to a polymerase chain reaction (PCR)? Select one:
hat componemts s hould be added to a polymerase chain reaction (PCR)? Select one:
at what temperature does the binding annealing of primers occour
at what temperature does the binding annealing of primers occour
An variant ofthe ACE gene allele with Alu insertion was identified:
An variant ofthe ACE gene allele with Alu insertion was identified:
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Study Notes
PCR Reaction
- Taq polymerase is the enzyme needed for the synthesis of a DNA strand.
- Primer hybridization occurs at approximately 50-65°C.
- Deoxyribonucleoside triphosphates (dNTPs) are missing from the PCR reaction mixture.
- Stages of PCR reaction:
- Denaturation: DNA is heated to separate the strands.
- Annealing: Primers bind to complementary sequences on the DNA strands.
- Extension: Taq polymerase extends the primers, synthesizing new DNA strands.
- Alu sequence is about 300 base pairs long.
- Electrophoresis takes approximately 30-60 minutes to detect the I and D alleles of the ACE gene.
- DNA fragment size:
- Smaller fragments migrate farther in the gel.
- Components of PCR reaction:
- DNA template: The DNA to be amplified.
- Primers: Short sequences of DNA that bind to the template DNA.
- Taq polymerase: An enzyme that synthesizes new DNA strands.
- dNTPs: Building blocks for the new DNA strands.
- PCR buffer: Provides optimal conditions for the reaction.
- Magnesium chloride (MgCl2): A cofactor for Taq polymerase.
- Primer annealing typically occurs at 50-65°C.
- Electrophoresis is the separation of DNA fragments based on size.
- Alu insertion is a variation in the ACE gene that can be identified by PCR.
ACE Gene
- People with higher levels of the ACE enzyme in their blood are likely to have the DD genotype.
- Hardy-Weinberg's law predicts the following ratio of genotypes for the ACE gene:
- DD: 0.64
- ID: 0.32
- II: 0.04
- PCR reaction is used to identify the ACE gene with Alu insertion.
Other Information
- Gel electrophoresis separates DNA fragments based on size, with smaller fragments migrating farther.
- DNA fragment amplified by ACE gene allele with a deletion is approximately 190 base pairs long.
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