Amplification Flashcards
What step within a cycle of PCR is related to the primers binding to the template DNA?
Annealing
1) What size range in bases are the repetitive sequences found in STRs?
2) What size range in bases are the repetitive sequences found in the PowerPlex Fusion 6C Amplification Kit?
1) 2-6 bases
2) 3-5 bases
What PCR cycle step does the DNA polymerase copy the template DNA?
Extension
What is the term for the separation of double stranded DNA in PCR?
Denaturation
Why is Taq DNA polymerase used in PCR over other DNA polymerase enzymes?
Thermostability
What is the total number of target genetic markers (autosomal and sex) in the PowerPlex Fusion 6C Amplification Kit?
27
(True or False) Magnesium chloride is needed for primer annealing but does not have any other functions in PCR reactions.
False; has many other functions
In what year did Kary Mullis invent PCR?
1983
In what year did Kary Mullis first publish about PCR?
1985
In what year did Kary Mullis receive the Nobel Prize for the invention of PCR?
1993
What is the most commonly used size of STR repeat units?
4 bases
What is the function of potassium chloride in a PCR reaction?
Neutralize the charge present on the backbone of DNA
Facilitate the annealing of the primer to the template DNA
Stabilize the primer template binding
What organism was Taq DNA polymerase originally isolated from?
Thermus Aquaticus
What are primers?
Chemically synthesized oglionucleotide fragments that flank the desired DNA segment to be amplified.
What number of fluorescent dyes are used for analysis in the PowerPlex Fusion 6c Amplification kit?
6
What is the most common method of dye labeling STR fragments in forensic DNA analysis?
Dye-labeled primers
(True or False) The desired PCR product is often called amplicon.
True
Why is there a hold step after PCR cycling?
To complete adenylation
(True or False) The addition of bovine serum albumin to PCR reactions reduces amplification inhibition.
True
The theoretical process of PCR results in a/an ______ accumulation of the target fragment beginning within the ______ cycle.
Exponential
3rd
(True or False) Thermal cyclers are instruments that have the ability to rapidly heat and cool which is why they were developed for PCR reactions.
True
Does Taq Polymerase add nucleotides to the 3’ or 5’ end of the DNA strand during extension?
3’
What are the 3 purposes of STR amplification?
- Target specific regions of DNA
- Generate millions of copies of target regions
- To detect STR fragments during capillary electrophoresis
What are STR markers?
Short DNA sequences that are repeated several times in a row
What is PCR?
Polymerase Chain Reaction
What are the PCR components necessary for amplification?
-DNA (template DNA)
-Primers
-Deoxynucleoside Triphosphates (dNTP’s)
-DNA polyermase
-Bovine Serum Albumin (BSA)
-Magnesium Chloride
-Potassium Chloried
