Term
The following are all types of what?
Reverse Transcriptase
Multiplex
Real Time |
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Definition
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Term
| What is the purpose of Reverse Transcriptase(3)? |
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Definition
Detect RNA in patient specimens
Detect RNA viruses
Determine gene "expression" |
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Term
True or False:
Sometimes mRNA is better than DNA for diagnosing disease |
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Definition
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Term
| Where is Reverse Transcriptase isolated from? |
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Definition
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Term
| What does Reverse Transcriptase do? |
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Definition
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Term
| "New" DNA (RNA --> DNA) = |
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Definition
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Term
What are the following considered:
HIV
HCV |
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Definition
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Term
| Why does Reverse Transcriptase use poly(T) primer? |
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Definition
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Term
True or False:
Reverse Transcriptase - All mRNAs will be present as single stranded cDNAs |
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Definition
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Term
Which of the following does NOT have introns:
Genomic
cDNA |
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Definition
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Term
Which of the following has BOTH introns and exons:
Genomic
cDNA |
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Definition
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Term
True or False:
Reverse transcription of Viral RNA into cDNA has different type of primers |
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Definition
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Term
| What kind of primers does Reverse transcription of Viral RNA use? |
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Definition
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Term
| Multiplex PCR - Amplification of how many target sequences in one reaction? |
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Definition
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Term
True or False:
Multiplex PCR need primer set for EACH target |
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Definition
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Term
| What is the purpose of Multiplex PCR? |
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Definition
| ID species & at same time detect target sequence |
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Term
What is the following an example of?:
Methicillin-resistant Staphylococcus aureus (MRSA) |
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Definition
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Term
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Definition
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Term
| Is Traditional PCR quantitative or semi-quantitative? |
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Definition
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Term
| How does Traditional PCR distinguish amplicons? |
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Definition
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Term
| What does Traditional PCR require? |
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Definition
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Term
| What does Real-Time PCR use? |
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Definition
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Term
| What does Real-Time PCR detect? |
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Definition
| PCR amplification during EARLY phase of Reaction |
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Term
| Is Real-Time PCR quantitative or semi-quantitative? |
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Definition
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Term
| What are the 3 phases of PCR? |
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Definition
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Term
| What happens during the Exponential stage? |
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Definition
Exact doubling of product every cycle
Reaction specific & precise |
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Term
| What happens during the Linear stage? |
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Definition
Reaction slowing
Products starting to degrade
Reactions components are being consumed |
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Term
| What happens during the Plateau stage? |
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Definition
Reaction has stopped
No more products are being made
End-point for gel detection |
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Term
| What phase is used in Real-Time PCR? |
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Definition
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Term
| Does the linear phase have a high variability? |
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Definition
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Term
True or False:
In the Plateau phase PCR products will degrade if left for too long |
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Definition
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Term
| What type of probe does the TaqMan Method use? |
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Definition
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Term
| What is Cycle Threshold (CT)? |
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Definition
| Number of PCR cycles to reach threshold where fluorescence detected |
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Term
| High initial concentration needs ____ cycles to reach CT |
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Definition
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Term
| Low initial concentration needs ____ cycles to reach CT |
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Definition
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Term
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Definition
| One dye (Q) interferes with fluorescence of the other (R) |
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Term
| What are the 2 types of flurophores in the TaqMan Method? |
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Definition
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Term
| What are the steps to the TaqMan Method? |
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Definition
1. Target DNA denatured to single strands by Heat
2. Primer binds to template/target DNA
3. TaqMan Probe binds to template/target DNA
4. Taq polymerase synthesizes new DNA |
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Term
| When the Taq polymerase reaches TaqMan Probe what happens? |
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Definition
| Removes Reporter & then entire TaqMan Probe including Quencher |
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Term
| In the TaqMan Method what can fluoresce? |
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Definition
| Reporter, because it is now free of the Quencher |
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Term
| What type of probe is used in the Molecular Beacon Method? |
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Definition
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Term
True or False:
Molecular Beacon Method is FRET based technology |
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Definition
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Term
| What are the steps when newly synthesized PCR products are denatured by heat in the Molecular Beacon Method? |
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Definition
1. Beacon molecule "ends" also denature
2. Target specific nucleotides attach to specific target during annealing phase |
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Term
| What is the Molecular beacon |
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Definition
*Short strands of single stranded DNA
*Nucleotide sequence of beacon customized for specific target |
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Term
| What type of configuration does Beacon form? |
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Definition
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Term
True or False:
At each end of the Beacon there is either R or Q |
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Definition
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Term
True or False:
In the Molecular Beacon Method, the Quencher dye interferes with the Reporter |
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Definition
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Term
| What part of the Beacon contains the target specific nucleotides? |
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Definition
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Term
| What are the steps when beacon hybridizes to target in the Molecular Beacon Method? |
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Definition
1. R & Q separated by disance
2. R can now fluoresce
3. R & Q still attached to nucleotide chain |
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Term
| As PCR product accumulates, what increases? |
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Definition
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Term
| Why type of probe does the SYBR Green Method use? |
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Definition
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Term
| What does the SYBR Green probe bind to? |
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Definition
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Term
| What type of binding does SYBR Green use? |
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Definition
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Term
| With the SYBR method, fluorescence increases as PCR ______ the amount of DNA |
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Definition
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