Font size
WorksheetsReal-time PCR Training
Total questions: 11
Worksheet time: 6mins
The abbreviation of Real-Time PCR is
PCR
qPCR
RT-PCR
RT-qPCR
Polymerase Chain Reaction (PCR) allows scientists to amplify (make many copies of) a small amount of DNA using a repetitive process involving DNA polymerase and short DNA primers
Yes
No
Absolute Quantification Experiment:
Requires probe assay that consists of two allele-specific probes.
Requires a standard curve to determine the absolute quantity of a
target in samples.
Requires endogenous control to establish
relative expression levels that reflect the real biological changes
Relative Quantification Experiment:
Requires probe assay that consists of two allele-specific probes.
Requires a standard curve to determine the absolute quantity of a
target in samples.
Requires endogenous control to establish
relative expression levels that reflect the real biological changes
Genotyping Experiment:
Requires probe assay that consists of two allele-specific probes.
Requires a standard curve to determine the absolute quantity of a
target in samples.
Requires endogenous control to establish
relative expression levels that reflect the real biological changes
Threshold Cycle (Ct):
Is the level of signal that reflects statistically significant increase over the calculated baseline signal.
Is the cycle number at which the fluorescent signal of the reaction crosses the threshold.
refers to how well the PCR doubles the
amount of DNA during each cycle.
Is ideally close to -3.32.
Below are components of a PCR reaction (choose ALL that apply).
DNA Template
DNA Polymerase
Primers
Nucleotides
Below are the Cycling steps in a PCR (choose ALL that apply).
Denaturation
Annealing
Extension
Truncation
Advantage of Real-Time PCR over PCR.
Involve amplification of DNA sequence
Can accomodate End-Point analysis
Can provide Qualitative results
Can provide Quantitative results
Below are the Chemistries of Real-Time PCR (choose ALL that apply).
SYBR GREEN
Probe detection
Ethidium Bromide (EtBr)
SYBR Safe
Function of a Passive Referenec Dye in Real-Time PCR.
Normalize for non-PCR related fluctuations in fluorescence
(e.g., caused by pipetting errors).
Enhances amplification of DNA sequence.
Serve as the reporter dye for target gene in Real-Time PCR.
A double-stranded DNA binding dye that fluoresce when excited by light source.
