Lectures 1-2-3

Question 1

AAC, for a calibrator sample is always equal to…

Answer 1

Zero

Question 2

The reporter molecule on a Taq man probe is found where?

Answer 2

At the 5’ end of the probe

Question 3

A qPCR assay provides the following result:
Samples A has a Ct value of 22,
Sample B has a Ct value of 21,
Sample C has a Ct value of 26. Following the completion of the qPCR run, what has the lowest amount of starting amplicon?

Answer 3

Sample C has the lowest amount of starting amplicon at the beginning of the qPCR assay

Question 4

In a Real Time PCR run of two samples, Sample A has a Ct value of 20 while Sample B has a Ct value of 17. This suggests what?

Answer 4

Sample A has eight fold less target sequence than sample B

Question 5

The filter sets that are included in real time PCR instruments to ensure that the optimal wavelengths of light for sending to the fluorophore and for optimal signal detection are referred to as what?

Answer 5

The excitation and emission filter sets

Question 6

The threshold setting value for fluorescence usually equates with a value that what?

Answer 6

Ten SDs above the baseline value

Question 7

What are passive reference dyes such as ROX used for

Answer 7

Normalising pipetting errors and normalises machines “noise”

Question 8

What does Ct refer to in real time PCR

Answer 8

The cycle at which the threshold value of fluorescence is reached by a reaction

Question 9

True or false, fluorophore fluorescence via ROX and normalised samples Ct values compared to a calibrator refer to the same phenomenon

Answer 9

False

Question 10

True or false, In comparative PCR, precise copy number determination is necessary

Answer 10

False

Question 11

True or false
RT-PCR is another name for reverse transcriptase PCR

Answer 11

True

Question 12

For FAM reporter, Rn is determined by what?

Answer 12

Dividing reporter fluorescence value by ROX fluorescence value

Question 13

True or false, RT-PCR is the acronym for Real Time Polymerase Chain Reaction

Answer 13

False

Question 14

The passive reference dye used in most qCR instruments is what?

Answer 14

ROX

Question 15

True or false, RT-PCR and qPCR are synonymous terms that refer to real time PCR

Answer 15

False

Question 16

What is a housekeeping gene also known as?

Answer 16

Endogenous control

Question 17

How many Ct values separate two samples in which one has ten times more starting material than the over?

Answer 17

3.32

Question 18

True or false, All qPCR assays require the inclusion of a standard curve

Answer 18

False

Question 19

The reporter molecule in the TAQMAN system fluorescences when?

Answer 19

It fluorescences constantly irrespective of whether amplification is taking place or not

Question 20

True or false
Test Sample A contains eight times more target copies than Test Sample B prior to commencing our qPCR run. At the threshold level of fluorescence, Sample B will therefore contain eight times less of the amplicon than Sample A.

Answer 20

False

Question 21

In qPCR, two peaks observed following melt curve analysis indicate what?

Answer 21

Two regions of DNA have been amplified, what they are being dependent on their melt temperature.

Question 22

True or false. In PCR, threshold values are always generated automatically and will have a value set at ten standard deviations above the baseline value. T/F

Answer 22

False

Question 23

AARn is a value that determines the relative expression of a gene in a test assay monitoring expression levels over a defined timer period,
true or false

Answer 23

False

Question 24

Amplification detection in the NTC wells of a Real Time PCR study….

Answer 24

Invalidates the sample results

Question 25

A direct correlation exists between template copy number in the sample vial pre-PCR and the early cycles of the _____

Answer 25

Exponential phase

Question 26

The plateau phase is___?

Answer 26

Not important in qPCR assays