p.14
Fluorescence Detection in qPCR
What indicates that the reaction has reached the upper limit of detection in qPCR?
The plateau of the fluorescence signal.
p.13
Fluorescence Detection in qPCR
What does the exponential increase in fluorescence signal indicate?
It indicates that the amplicon is exponentially increasing in number.
p.41
Comparison of PCR and qPCR Techniques
What does RT-qPCR stand for?
Quantitative Reverse Transcription PCR.
p.14
Fluorescence Detection in qPCR
What happens to the fluorescence signal after several more cycles in qPCR?
The fluorescence signal plateaus.
p.35
Role of Reverse Transcriptase in RT-PCR
Why did researchers use RT-PCR to detect the Zika virus instead of standard PCR?
RT-PCR is needed to detect RNA viruses like Zika, while standard PCR is used for DNA.
p.35
Analyzing Ct Values and Standard Curves
Is the RT-PCR experiment intended to be quantitative?
No, it is primarily qualitative to confirm the presence of the Zika virus.
p.36
Role of Reverse Transcriptase in RT-PCR
What is the purpose of reverse transcription in RT-PCR?
To convert RNA into complementary DNA (cDNA) that can be used as a PCR template.
p.11
Fluorescence Detection in qPCR
What is the status of detection in the initial cycles of qPCR?
We are below the lower limit of detection of our instrument.
p.28
Experimental Design for Monitoring CAR T-cell Abundance
What is the purpose of the qPCR experiment in CAR T-cell therapy?
To monitor CAR T-cell abundance over time.
p.2
Copy Number Polymorphism (CNP) Implications
What are potential consequences of having more copies of a gene than normal?
May lead to gene dosage effects, altered gene expression, and potential health issues.
p.9
Fluorescence Detection in qPCR
When do molecules exhibit fluorescence?
When bound to double-stranded DNA at a specific wavelength.
p.5
Copy Number Polymorphism (CNP) Implications
What technique can be used to analyze the copy number of a gene within a person's genome?
Real-time PCR (also called quantitative PCR; qPCR).
p.41
Quantitative PCR (qPCR) Fundamentals
What is done after producing cDNA in RT-qPCR?
The cDNA is quantified using the qPCR process.
p.34
Virus Detection Using RT-PCR
In which type of samples was the Zika virus detected using RT-PCR?
Human blood serum samples.
p.2
Copy Number Polymorphism (CNP) Implications
What is Copy Number Polymorphism (CNP) in the human genome?
Variations in the number of copies of a particular gene.
p.2
Copy Number Polymorphism (CNP) Implications
How common is Copy Number Polymorphism (CNP) in the human genome?
More common than previously thought.
p.15
Analyzing Ct Values and Standard Curves
What does the qPCR machine record for each sample?
The cycle number at which the sample surpasses the fluorescence threshold.
p.28
Analyzing Ct Values and Standard Curves
What specific analysis is performed on the blood DNA in the qPCR experiment?
Analyzing the copy number of the transgene present in a specific amount of blood DNA.
p.29
Applications of RT-qPCR in Gene Expression Analysis
What are CAR T-cells?
T-cells that are isolated from a patient, genetically engineered with a therapeutic transgene, and reinfused back into the patient to fight specific types of cancer.
p.31
Role of Reverse Transcriptase in RT-PCR
What does viral reverse transcriptase do in retrovirus replication?
Copies (+)RNA into (-)DNA (cDNA).
p.6
Analyzing Ct Values and Standard Curves
When are amplicons assessed in qPCR?
In real-time, at the end of each cycle.
p.30
Role of Reverse Transcriptase in RT-PCR
Why is it challenging to clone a poliovirus gene directly?
Because poliovirus is an RNA virus and has no DNA to copy.
p.21
Analyzing Ct Values and Standard Curves
What is the purpose of analyzing DNA samples from two different patients in this example?
To examine whether there is copy number polymorphism of a specific gene of interest.
p.41
Role of Reverse Transcriptase in RT-PCR
What is the first step in the RT-qPCR process?
Reverse transcribing the RNA sample to produce cDNA.
p.11
Fluorescence Detection in qPCR
What happens to amplicon abundance in the first several cycles of qPCR?
Amplicon abundance is low.
p.4
Comparison of PCR and qPCR Techniques
What does Standard PCR + Agarose Gel Electrophoresis indicate about a gene?
It can only tell you whether the gene is present (yes or no).
p.7
Analyzing Ct Values and Standard Curves
What does the level of fluorescence in qPCR correlate with?
The amount of DNA in the sample.
p.31
Role of Reverse Transcriptase in RT-PCR
What is the first step in the replication of a retrovirus?
(+)ssRNA viral genome enters the host cell.
p.8
Fluorescence Detection in qPCR
Who conducted the study referenced regarding dye molecules and dsDNA?
Galindo-Murillo et al. in 2021.
p.26
Analyzing Ct Values and Standard Curves
How can a standard curve be generated for a qPCR experiment?
By performing serial 10-fold dilutions of a DNA sample containing a known copy number of the transgene of interest.
p.33
Quantitative PCR (qPCR) Fundamentals
What components are needed for PCR amplification after cDNA synthesis?
DNAP, primers, and dNTPs are needed to produce dsDNA.
p.31
Role of Reverse Transcriptase in RT-PCR
What does host DNA-dependent RNA polymerase transcribe?
(+)mRNA which gets translated into viral proteins.
p.38
Virus Detection Using RT-PCR
What should the negative serum sample have produced in the Zika virus detection study?
No band, confirming no viral RNA was present.
p.42
Applications of RT-qPCR in Gene Expression Analysis
How much is fabB up-regulated under low pH conditions?
~2.5-fold (2.5x higher expression).
p.36
Role of Reverse Transcriptase in RT-PCR
Why did researchers use RT-PCR instead of standard PCR for Zika virus detection?
To detect the Zika virus genome, which consists of single-stranded RNA that requires reverse transcription to produce cDNA for amplification.
p.37
Virus Detection Using RT-PCR
Is the RT-PCR method used in this experiment quantitative?
No, it is not sensitive enough to quantify Zika virus levels.
p.7
Fluorescence Detection in qPCR
When does the qPCR machine add a fluorescence detection step?
After the elongation/extension step of each cycle.
p.15
Fluorescence Detection in qPCR
What does the qPCR machine set to detect fluorescence increases?
An arbitrary fluorescence threshold.
p.28
Experimental Design for Monitoring CAR T-cell Abundance
What samples would researchers test using qPCR?
Blood samples from the same patients at different time points post-infusion with CAR T-cells.
p.20
Analyzing Ct Values and Standard Curves
What is the purpose of creating a standard curve in qPCR?
To quantify the amount of target DNA in experimental samples.
p.16
Analyzing Ct Values and Standard Curves
What is the first step in creating a standard curve for qPCR?
Start with a 'known' sample with a known amount of target DNA.
p.4
Comparison of PCR and qPCR Techniques
What is the limitation of using Standard PCR for gene analysis?
It cannot provide quantitative data on gene copy number.
p.16
Analyzing Ct Values and Standard Curves
How are dilutions made for qPCR standard curves?
By making 10-fold dilutions of the known sample.
p.22
Analyzing Ct Values and Standard Curves
If the gene of interest is on chromosome 20, what should be the relative concentration in both samples?
The relative concentration of chromosome 20 should be the same in both samples.
p.27
Experimental Design for Monitoring CAR T-cell Abundance
What is the purpose of a 'no template' control in qPCR experiments?
To assess and subtract the background noise generated by fluorescent molecules in the solution.
p.42
Applications of RT-qPCR in Gene Expression Analysis
What was the control condition in the RT-qPCR experiment?
Growth in neutral media (pH 7).
p.38
Virus Detection Using RT-PCR
What do the negative control results confirm in the Zika virus detection study?
That the approach specifically detected Zika virus, not RNA viruses broadly.
p.33
Quantitative PCR (qPCR) Fundamentals
What is the significance of the PCR cycle in RT-PCR?
The PCR cycle amplifies the cDNA to produce double-stranded DNA (dsDNA).
p.23
Analyzing Ct Values and Standard Curves
What could the copy numbers of the target gene imply for Patient 1 and Patient 2?
Patient 1 could have 4 copies and Patient 2 1 copy, or Patient 1 could have 8 copies and Patient 2 2 copies.
p.39
Role of Reverse Transcriptase in RT-PCR
What would be the expected result if reverse transcriptase is omitted from the RT-PCR reaction?
No amplification of Zika virus RNA would be expected.
p.13
Fluorescence Detection in qPCR
What happens to the fluorescence signal during the next several cycles of PCR?
The fluorescence signal increases exponentially.
p.12
Fluorescence Detection in qPCR
What is the significance of reaching 'critical mass' in qPCR?
It allows for the detection of the fluorescence signal.
p.7
Quantitative PCR (qPCR) Fundamentals
What does qPCR monitor during the PCR process?
The amplification of target DNA in real time.
p.35
Role of Reverse Transcriptase in RT-PCR
What result would you expect if reverse transcriptase was omitted from the reaction?
You would expect no amplification of the Zika virus RNA.
p.7
Fluorescence Detection in qPCR
What happens to the fluorophore in the reaction mixture during qPCR?
It is excited by a specific wavelength of light and emits a different wavelength of light.
p.4
Comparison of PCR and qPCR Techniques
Why is Standard PCR not suitable for quantifying gene copies?
It is not quantitative enough to tell you how many copies of the gene are present.
p.6
Comparison of PCR and qPCR Techniques
How does qPCR differ from traditional PCR?
qPCR uses fluorescent dyes or probes to detect DNA molecules.
p.42
Applications of RT-qPCR in Gene Expression Analysis
What does RT-qPCR measure in an experimental treatment sample?
The level of a specific mRNA relative to a control sample.
p.6
Quantitative PCR (qPCR) Fundamentals
What type of thermal cycler is required for qPCR?
A special thermal cycler with fluorescence detection capability.
p.40
Applications of RT-qPCR in Gene Expression Analysis
What environmental condition was tested for its effect on catalase activity?
Different pH levels of the media.
p.18
Analyzing Ct Values and Standard Curves
Why does each dilution in PCR start with 10x fewer template DNA molecules?
Because we made 10-fold dilutions.
p.29
Analyzing Ct Values and Standard Curves
What was the result for Patient 2 regarding CAR T-cells?
No CAR T-cells were detected in the blood at any timepoint.
p.33
Role of Reverse Transcriptase in RT-PCR
What is the sequence of the cDNA template provided?
C T C T A A G G T C A 5’ 3’.
p.37
Virus Detection Using RT-PCR
What type of primers were used in the RT-PCR experiment?
Primers specific to the Zika virus.
p.5
Comparison of PCR and qPCR Techniques
What is the difference between PCR and qPCR?
qPCR quantifies the amount of target DNA, while traditional PCR only amplifies it.
p.17
Analyzing Ct Values and Standard Curves
What does a perfectly set up standard curve indicate in PCR?
It indicates that each subsequent dilution requires 3.2 additional cycles to surpass the fluorescence threshold.
p.36
Virus Detection Using RT-PCR
What does RT-PCR stand for?
Reverse Transcription Polymerase Chain Reaction.
p.8
Fluorescence Detection in qPCR
What do dye molecules do to dsDNA?
They intercalate, stacking between base pairs.
p.17
Analyzing Ct Values and Standard Curves
What does the term 'fluorescence threshold' refer to in PCR?
The level of fluorescence at which the PCR product can be reliably detected.
p.22
Analyzing Ct Values and Standard Curves
What is another important assumption made when comparing the two patient samples?
The concentration of DNA is the same for both patient samples.
p.1
Role of Reverse Transcriptase in RT-PCR
What is the role of reverse transcriptase in RT-PCR?
To convert RNA into complementary DNA (cDNA).
p.30
Role of Reverse Transcriptase in RT-PCR
What is the purpose of using Reverse Transcription PCR (RT-PCR) in this scenario?
To clone a poliovirus gene into a vector for protein expression in E. coli.
p.1
Applications of RT-qPCR in Gene Expression Analysis
What is a key aspect of interpreting RT-PCR and RT-qPCR results?
Understanding the amplification curves and Ct values.
p.19
Analyzing Ct Values and Standard Curves
What is the fluorescence threshold in PCR?
The point at which the fluorescence signal becomes detectable above the background noise.
p.43
Differentiating Between PCR Techniques
What is Standard PCR used to determine?
Whether a specific gene or organism is present in a sample (yes/no).
p.43
Differentiating Between PCR Techniques
What does RT-PCR help to identify?
Whether a specific RNA virus is present in a sample (yes/no).
p.12
Fluorescence Detection in qPCR
What happens to amplicon density after several cycles in qPCR?
It increases beyond the 'critical mass' required to detect the fluorescence signal.
p.37
Virus Detection Using RT-PCR
What does the agarose gel in the experiment show?
The results of RT-PCR assessment of human serum samples for Zika virus.
p.35
Comparison of PCR and qPCR Techniques
What do the results of negative controls with serum samples from patients with other infections confirm?
They confirm the specificity of the RT-PCR for the Zika virus.
p.34
Virus Detection Using RT-PCR
In which publication was the study on Zika virus detection using RT-PCR published?
Memórias do Instituto Oswaldo Cruz.
p.20
Analyzing Ct Values and Standard Curves
How is the Ct value of an experimental sample used in qPCR?
It is compared to the standard curve to quantify the amount of target DNA.
p.6
Quantitative PCR (qPCR) Fundamentals
What does qPCR monitor during the PCR process?
The amplification of target DNA in real time.
p.18
Analyzing Ct Values and Standard Curves
What is the relationship between PCR cycles and the number of amplicons?
The number of amplicons doubles after every cycle (2^n, where n = number of PCR cycles).
p.19
Analyzing Ct Values and Standard Curves
If Sample 1 contains 100 template molecules, how many template molecules does Sample 2 contain?
10 template molecules (a 10-fold dilution of Sample 1).
p.29
Analyzing Ct Values and Standard Curves
What was observed in Patients 1 and 3 regarding CAR T-cell abundance from days 7-14 post-infusion?
A slight increase in copy number, indicating a higher number of CAR T-cells.
p.1
Comparison of PCR and qPCR Techniques
What should be determined to solve different scientific problems using PCR?
The best PCR-based approach.
p.43
Differentiating Between PCR Techniques
When should you use Standard PCR?
When you need a large amount of DNA for cloning, sequencing, or mutational analysis.
p.23
Analyzing Ct Values and Standard Curves
What is the ratio of target gene copies between Patient 1 and Patient 2?
Patient 1 has approximately 4 times more copies than Patient 2.
p.43
Differentiating Between PCR Techniques
When is RT-PCR used?
When you need to amplify DNA from RNA for cloning, sequencing, or mutational analysis.
p.32
Role of Reverse Transcriptase in RT-PCR
What is the role of reverse transcriptases in RT-PCR?
They convert RNA into complementary DNA (cDNA).
p.10
Fluorescence Detection in qPCR
What activity does DNAP have during the extension phase?
5’ to 3’ exonuclease activity, degrading the probe.
p.5
Quantitative PCR (qPCR) Fundamentals
What does qPCR measure in a sample?
How much of the target gene is present.
p.15
Fluorescence Detection in qPCR
What performs the analysis of qPCR fluorescence curves?
The qPCR machine and its companion software.
p.16
Analyzing Ct Values and Standard Curves
What does the standard curve represent in qPCR analysis?
It represents our positive control and measurement standard.
p.26
Experimental Design for Monitoring CAR T-cell Abundance
What are CAR T-cells?
T-cells that are isolated from a patient, genetically engineered with a therapeutic transgene, and reinfused back into the patient to fight specific types of cancer.
p.38
Virus Detection Using RT-PCR
What were the negative controls used in the Zika virus detection study?
Serum samples from patients with active infections of Chikungunya virus (CHIKV), Dengue virus (DENV), or negative for any infection.
p.26
Analyzing Ct Values and Standard Curves
What is the required initial copy number for the undiluted sample in the qPCR experiment?
More than 100,000 copies.
p.29
Analyzing Ct Values and Standard Curves
What happened to CAR T-cell levels in Patients 1 and 3 after 28-56 days?
There was a rapid decrease to 0.
p.40
Applications of RT-qPCR in Gene Expression Analysis
What trend is observed in catalase activity as pH decreases?
Catalase activity increases as pH decreases.
p.39
Virus Detection Using RT-PCR
What does the agarose gel show in the context of Zika virus detection?
Results of RT-PCR assessment of human serum samples.
p.25
Experimental Design for Monitoring CAR T-cell Abundance
What are CAR T-cells?
T-cells that are isolated from a patient, genetically engineered with a therapeutic transgene, and reinfused back into the patient to fight specific types of cancer.
p.43
Differentiating Between PCR Techniques
What can RT-qPCR quantify?
Relative gene expression (mRNA) or the number of RNA viruses present in a sample.
p.28
Applications of RT-qPCR in Gene Expression Analysis
What are CAR T-cells?
T-cells that are isolated from a patient, genetically engineered with a therapeutic transgene, and reinfused back into the patient to fight specific types of cancer.
p.16
Analyzing Ct Values and Standard Curves
What is the purpose of comparing the Ct value of an experimental sample?
To compare it to a standard curve.
p.17
Analyzing Ct Values and Standard Curves
What is the significance of surpassing the fluorescence threshold in PCR?
It indicates that the target DNA has been amplified sufficiently for detection.
p.40
Applications of RT-qPCR in Gene Expression Analysis
What hypothesis did the labmate propose regarding the katA gene?
The katA gene is up-regulated when P. enteraalne is exposed to low pH.
p.40
Applications of RT-qPCR in Gene Expression Analysis
What does gene expression analysis measure?
The amount of mRNA being produced for a specific gene.
p.26
Experimental Design for Monitoring CAR T-cell Abundance
Why do researchers need to monitor CAR T-cell abundance?
To understand how long the CAR T-cells persist in the body.
p.40
Applications of RT-qPCR in Gene Expression Analysis
How does mRNA level relate to protein production?
More mRNA indicates more of that protein being produced.
p.42
Applications of RT-qPCR in Gene Expression Analysis
What was the experimental treatment condition for the RT-qPCR experiment?
Growth in acidic media (pH 4.2).
p.38
Virus Detection Using RT-PCR
What does a negative result in the serum sample confirm about the RT-PCR setup?
That the RT-PCR setup was working properly.
p.25
Experimental Design for Monitoring CAR T-cell Abundance
What samples would you test using qPCR?
Blood samples from the patient at various time points post-infusion.
p.3
Quantitative PCR (qPCR) Fundamentals
What process occurs during transcription?
The synthesis of mRNA from a DNA template.
p.37
Virus Detection Using RT-PCR
What type of results does this RT-PCR method provide?
A yes/no answer (samples are either positive or negative for Zika virus).
p.20
Analyzing Ct Values and Standard Curves
What does the qPCR software do with the data points for each standard?
It condenses them into a standard curve.
p.22
Analyzing Ct Values and Standard Curves
What is the basis for comparing Patient 1 and Patient 2 samples in qPCR analysis?
The comparison is based on the assumption that the amount of input DNA is the same for both samples.
p.19
Analyzing Ct Values and Standard Curves
What does a perfectly set up standard curve indicate in PCR?
It takes 3.2 additional cycles of PCR for each subsequent dilution to surpass the fluorescence threshold.
p.29
Experimental Design for Monitoring CAR T-cell Abundance
What is the purpose of the qPCR experiment in the context of CAR T-cells?
To monitor CAR T-cell abundance over time.
p.33
Role of Reverse Transcriptase in RT-PCR
What is the purpose of complementary DNA (cDNA) in RT-PCR?
cDNA is used as a template for PCR amplification.
p.30
Role of Reverse Transcriptase in RT-PCR
How does Reverse Transcriptase defy the Central Dogma?
By synthesizing DNA from RNA.
p.30
Role of Reverse Transcriptase in RT-PCR
What is the ultimate goal of cloning the poliovirus gene?
To express the protein in E. coli for further analysis.
p.10
Fluorescence Detection in qPCR
What happens to the fluorophore when the probe is degraded?
The fluorophore is released and is no longer inhibited by the quencher.
p.25
Comparison of PCR and qPCR Techniques
What is a good negative control for the qPCR experiment?
A sample that does not contain CAR T-cells or a non-target gene.
p.24
Analyzing Ct Values and Standard Curves
What could the copy numbers of the target gene imply for Patient 1 and Patient 2?
Patient 1 could have 4 copies and Patient 2 1 copy, or Patient 1 could have 8 copies and Patient 2 2 copies.
p.3
Applications of RT-qPCR in Gene Expression Analysis
How does gene copy number affect gene dosage?
It influences how much mRNA and therefore protein is being synthesized.
p.27
Experimental Design for Monitoring CAR T-cell Abundance
What are CAR T-cells?
T-cells that are isolated from a patient, genetically engineered with a therapeutic transgene, and reinfused back into the patient to fight specific types of cancer.
p.1
Analyzing Ct Values and Standard Curves
What does the Ct value correlate with?
Target DNA copy number, given appropriate standards.
p.27
Experimental Design for Monitoring CAR T-cell Abundance
Why do researchers need to monitor CAR T-cell abundance?
To understand how long the CAR T-cells persist in the body.
p.23
Analyzing Ct Values and Standard Curves
What can be determined from analyzing qPCR fluorescence curves in patients?
The relative number of copies of the gene target present in their genomes.
p.38
Virus Detection Using RT-PCR
What does the agarose gel show in the context of Zika virus detection?
Results of RT-PCR assessment of human serum samples specific to the Zika virus.
p.31
Role of Reverse Transcriptase in RT-PCR
What happens to the RNA strand during retrovirus replication?
The RNA strand is degraded.
p.19
Analyzing Ct Values and Standard Curves
What is the relationship between template concentration and PCR cycles in a standard curve?
Each 10-fold dilution requires 3.2 additional cycles to surpass the fluorescence threshold.
p.21
Analyzing Ct Values and Standard Curves
What was the initial standard copy number of the target gene?
Log 10 = 7.5 (10^7.5 copies).
p.31
Role of Reverse Transcriptase in RT-PCR
What is produced from the copies made by host DNA-dependent RNA polymerase?
(+)ssRNA genome, which are packaged into new virions.
p.43
Differentiating Between PCR Techniques
What is the purpose of RT-qPCR?
To quantify how much of a specific RNA target is in your sample.
p.25
Experimental Design for Monitoring CAR T-cell Abundance
Why do researchers need to monitor CAR T-cell abundance?
To understand how long the CAR T-cells persist in the body.
p.43
Differentiating Between PCR Techniques
What can qPCR quantify?
Gene copy number, plasmid copy number, number of bacteria, or DNA viruses present in a sample.
p.18
Analyzing Ct Values and Standard Curves
What is the formula used to calculate the number of additional cycles required for a 10-fold dilution to reach the same threshold?
2^n = 10, which gives n = √10 = 3.2.
p.42
Applications of RT-qPCR in Gene Expression Analysis
How much is fabA up-regulated under low pH conditions?
~2-fold (2x higher expression).
p.10
Fluorescence Detection in qPCR
What is the role of hybridization probes in qPCR?
They offer more specific detection of a particular amplicon.
p.10
Fluorescence Detection in qPCR
What are hybridization probes linked to?
Both a fluorophore and a quencher.
p.39
Role of Reverse Transcriptase in RT-PCR
What does the presence of amplification indicate if reverse transcriptase is omitted?
It would indicate contamination or the presence of DNA, not RNA.
p.39
Role of Reverse Transcriptase in RT-PCR
Why is reverse transcriptase important in RT-PCR?
It converts RNA into complementary DNA (cDNA) for amplification.
p.25
Analyzing Ct Values and Standard Curves
How would you generate a standard curve for the qPCR experiment?
By using known concentrations of CAR T-cell DNA to create a plot of Ct values against the log of the initial quantity.
p.24
Analyzing Ct Values and Standard Curves
What can be determined from analyzing qPCR fluorescence curves in patients?
The relative number of copies of the gene target present in their genomes.
p.3
Quantitative PCR (qPCR) Fundamentals
What is the role of translation in the Central Dogma?
It is the process where mRNA is used to synthesize proteins.
p.32
Role of Reverse Transcriptase in RT-PCR
What is the significance of cDNA in molecular biology?
It serves as a template for PCR amplification.
p.10
Fluorescence Detection in qPCR
What does the quencher do in the presence of the fluorophore?
It inhibits fluorescence when they are in close proximity.
p.43
Differentiating Between PCR Techniques
What does qPCR measure?
How much of a specific DNA target is in your sample.
p.3
Quantitative PCR (qPCR) Fundamentals
What is the Central Dogma of molecular biology?
It describes the flow of genetic information from DNA to RNA (transcription) and then to protein (translation).
p.32
Role of Reverse Transcriptase in RT-PCR
In which organisms are reverse transcriptases found?
Retroviruses, retrotransposons, and eukaryotes (e.g., telomerase).
p.10
Fluorescence Detection in qPCR
How does the level of fluorescence relate to amplicon synthesis?
It correlates with the number of amplicons that have been synthesized.
p.25
Analyzing Ct Values and Standard Curves
How would you interpret the data from the qPCR experiment?
By analyzing the Ct values to determine the abundance of CAR T-cells over time, indicating their persistence in the body.
p.32
Role of Reverse Transcriptase in RT-PCR
What is produced at the end of the RT-PCR process?
Complementary DNA (cDNA).
p.24
Analyzing Ct Values and Standard Curves
What is the ratio of target gene copies between Patient 1 and Patient 2?
Patient 1 has approximately 4 times more copies than Patient 2.
