Term
| List the 7 steps of the Hybrid Capture Assay |
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Definition
1. Denature target DNA
2. SS-DNA + RNA Probe -> RNA-DNA Hybrid
3. RNA-DNA hybrid binds to anti-hybrid ABYs attached to tube
4. 2nd ABY (w/enzyme) added
5. 2nd Anti-hybrid ABY binds to hybrid
6. Add substrate
7. Measure amount of light |
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Term
Hybrid Capture Assay:
Enzyme + Substrate --> ? |
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Definition
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Term
| What does the Hybrid Capture Assay amplify? |
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Definition
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Term
| List the 6 steps of the Branched-chain DNA Assay |
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Definition
1. Specimen treated
2. Tube has multiple target-specific probes
3. Sample RNA binds to these capture probes
4. Amplification occurs as several sets of probes bind to target &/or to each other
5. Substrate added
6. Measure amount of light |
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Term
| What does the Branched-chain DNA Assay amplify? |
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Definition
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Term
Branched-chain DNA Assay:
Target + Capture Probe --> ? |
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Definition
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Term
Branched-chain DNA Assay:
Enzyme Substrate --> ? |
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Definition
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Term
| What happens in the Branched-chain DNA Assay when the specimen is treated? |
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Definition
Cells lysed
RNases inactivated |
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Term
| What are the 3 probes with Branched-chain DNA Assay? |
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Definition
Extender
Preamplifier
Amplifier |
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Term
| With the Branched-chain DNA Assay what do you use the series of oligonucleotide probes to do? |
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Definition
Immobilize target to tube/well
Amplify chemiluminescent signal through network of overlapping sequences to form a branched complex |
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Term
| By amplifying the chemiluminescent signal through network of overlapping sequences what do you form? |
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Definition
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Term
| What does the Cleavase-Invader Technology amplify? |
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Definition
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Term
| What are the 4 components of the Cleavase-Invader Technology? |
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Definition
Target DNA
Oligonucleotides
Cleavase
FRET Casette |
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Term
| What are the 2 Oligonucleotides that are part of the Cleavase-Invader Technology? |
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Definition
Primary Probe
Invader Oligo |
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Term
Which of the following is specific for the target?:
Primary Probe
Invader Oligo |
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Definition
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Term
| Which component of the Cleavase-Invader Technology cleaves "overlapping flaps"? |
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Definition
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Term
| What type of structure is the FRET Casette? |
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Definition
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Term
True or False:
In the Cleavase-Invader Technology the Fluorophore and Quencher are in close proximity |
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Definition
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Term
| What is the Primary Reaction of the Cleavase-Invader Technology? |
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Definition
| Primary Probe & Invader Oligo bind to target in an overlapping manner |
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Term
| The part of the Primary probe not binding to target = ? |
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Definition
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Term
| When Cleavase removes the Flap what does it become? |
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Definition
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Term
| What is the Secondary Reaction of the Cleavase-Invader Technology? |
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Definition
1.Cleaved Flap attaches to FRET Casette & Forms another flap
2. Cleavase removes casette flap and releases Reporter |
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Term
| After Cleavase removes the Casette Flap what can now fluoresce? |
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Definition
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Term
| Where does Amplification occur with the Cleavase-Invader Technology(2)? |
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Definition
*Attachment of Primary & Invader probes to target "cycles"
*Cleaved Flap |
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Term
True or False:
Flap formed each time Primary probe & Oligo probe attach to target |
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Definition
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Term
| What releases each Flap in Cleavase-Invader Technology? |
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Definition
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Term
True or False:
Cleavase-Invader Technology - 1 Target can cause release of Multiple Flaps |
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Definition
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Term
| What leads to the release of Reporter and Increased Fluorescence? |
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Definition
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Term
True or False:
Cleavase-Invader Technology can detect single point mutations |
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Definition
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Term
True or False:
Cleavase-Invader Technology is NOT as versatile as PCR |
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Definition
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