Quantification

Question 1

(True or False) The human male (Y) target is single copy to reduce variability between individuals and to correlate with STR amplicons.

Answer 1

False; the human male target is multicopy

Question 2

When using the QuantStudio instrument, light passes through five ______ and the ______ adhesive cover before reaching the samples in the ______.

Answer 2

Excitation Filters
Optical
Wells

Question 3

What is the name of the chemistry that is used in many quant kits?

Answer 3

TaqMan Chemistry

Question 4

What organism was Taq DNA polymerase originally isolated from?

Answer 4

Thermus Aquaticus

Question 5

What is the amplicon length of the human (autosomal) target for the PowerQuant System?

Answer 5

84 bases

Question 6

What is the purpose of a passive reference dye in qPCR?

Answer 6

Normalization

Question 7

What is the amplicon length(s) of the human male (Y) target for the PowerQuant System?

Answer 7

81 bases and 136 bases

Question 8

What qPCR phase shows that no more product is being produced?

Answer 8

Plateau

Question 9

What volume of sample, control, and DNA standards are added to a PowerQuant reaction?

Answer 9

2 microliters

Question 10

What is the phase of qPCR that shows a slowing of the amplification efficiency?

Answer 10

Linear

Question 11

What is the reaction volume of a PowerQuant reaction?

Answer 11

20 microliters

Question 12

What is the point in which fluorescence passes the baseline noise in qPCR?

Answer 12

Cycle Threshold or Quantification Cycle

Question 13

Why is quantification used in forensic DNA testing?

Answer 13

To determine the quantity and quality of human DNA

Question 14

A sample with inhibition would have a/an ______ IPC CT value

Answer 14

Elevated

Question 15

What is the measure of the closeness of fit between the standard curve regression line and the individual CT data points?

Answer 15

R squared value

Question 16

(True or False) The TaqMan chemistry uses probes with a reporter dye and a quencher that anneal within a target sequence outside forward and reverse primers.

Answer 16

False; between forward and reverse primers

Question 17

The CT value is ______ proportional to the ______ of the DNA sample.

Answer 17

Inversely
Concentration

Question 18

What is the passive reference dye for PowerQuant?

Answer 18

CXR

Question 19

What is a feature of modern qPCR chemistries that can show inhibition within a sample?

Answer 19

Internal PCR Control (IPC)

Question 20

What must be prepared and added to a qPCR run in order to determine the concentration of a DNA sample?

Answer 20

DNA Standard Dilution Series

Question 21

Which qPCR phase is related to the doubling of amplicons in each cycle at 100% efficiency?

Answer 21

Exponential

Question 22

What slope indicates 100% PCR amplification efficiency?

Answer 22

-3.3

Question 23

What is the stock concentration of the PowerQuant standard?

Answer 23

50 nanogram/microliter

Question 24

(True or False) The human (autosomal) target is multicopy to reduce variability between individuals and to correlate with STR amplicons.

Answer 24

True

Question 25

How many standard dilution points should be created when using the PowerQuant System at LSPCL?

Answer 25

4

Question 26

What does the slope of the standard curve indicate?

Answer 26

PCR efficiency

Question 27

What does the y-intercept of the standard curve indicate?

Answer 27

It is the expected ct/cq value for a sample at 1ng

Question 28

Does the PowerQuant System measure an increase or decrease in fluorescence?

Answer 28

Increase in fluorescence