Amplification

Question 1

What step within a cycle of PCR is related to the primers binding to the template DNA?

Answer 1

Annealing

Question 2

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?

Answer 2

1) 2-6 bases
2) 3-5 bases

Question 3

What PCR cycle step does the DNA polymerase copy the template DNA?

Answer 3

Extension

Question 4

What is the term for the separation of double stranded DNA in PCR?

Answer 4

Denaturation

Question 5

Why is Taq DNA polymerase used in PCR over other DNA polymerase enzymes?

Answer 5

Thermostability

Question 6

What is the total number of target genetic markers (autosomal and sex) in the PowerPlex Fusion 6C Amplification Kit?

Answer 6

27

Question 7

(True or False) Magnesium chloride is needed for primer annealing but does not have any other functions in PCR reactions.

Answer 7

False; has many other functions

Question 8

In what year did Kary Mullis invent PCR?

Answer 8

1983

Question 9

In what year did Kary Mullis first publish about PCR?

Answer 9

1985

Question 10

In what year did Kary Mullis receive the Nobel Prize for the invention of PCR?

Answer 10

1993

Question 11

What is the most commonly used size of STR repeat units?

Answer 11

4 bases

Question 12

What is the function of potassium chloride in a PCR reaction?

Answer 12

Neutralize the charge present on the backbone of DNA
Facilitate the annealing of the primer to the template DNA
Stabilize the primer template binding

Question 13

What organism was Taq DNA polymerase originally isolated from?

Answer 13

Thermus Aquaticus

Question 14

What are primers?

Answer 14

Chemically synthesized oglionucleotide fragments that flank the desired DNA segment to be amplified.

Question 15

What number of fluorescent dyes are used for analysis in the PowerPlex Fusion 6c Amplification kit?

Answer 15

6

Question 16

What is the most common method of dye labeling STR fragments in forensic DNA analysis?

Answer 16

Dye-labeled primers

Question 17

(True or False) The desired PCR product is often called amplicon.

Answer 17

True

Question 18

Why is there a hold step after PCR cycling?

Answer 18

To complete adenylation

Question 19

(True or False) The addition of bovine serum albumin to PCR reactions reduces amplification inhibition.

Answer 19

True

Question 20

The theoretical process of PCR results in a/an ______ accumulation of the target fragment beginning within the ______ cycle.

Answer 20

Exponential
3rd

Question 21

(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.

Answer 21

True

Question 22

Does Taq Polymerase add nucleotides to the 3’ or 5’ end of the DNA strand during extension?

Answer 22

3’

Question 23

What are the 3 purposes of STR amplification?

Answer 23

1. Target specific regions of DNA
2. Generate millions of copies of target regions
3. To detect STR fragments during capillary electrophoresis

Question 24

What are STR markers?

Answer 24

Short DNA sequences that are repeated several times in a row

Question 25

What is PCR?

Answer 25

Polymerase Chain Reaction

Question 26

What are the PCR components necessary for amplification?

Answer 26

-DNA (template DNA)
-Primers
-Deoxynucleoside Triphosphates (dNTP’s)
-DNA polyermase
-Bovine Serum Albumin (BSA)
-Magnesium Chloride
-Potassium Chloried