Term
| Billions of copies can be made from how many copies of target DNA? |
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Definition
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Term
What are the following considered:
Detect MOs
Detect Gene Defect
Forensics
Transplants (HLA typing) |
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Definition
| Applications of PCR & PCR variations |
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Term
| What are the components needed for PCR? |
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Definition
DNA target
2 Primers
Taq DNA polymerase
Deoxynucleotide triphosphates
Buffer with Mg+2 |
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Term
| Why do you use a buffer with Magnesium +2? |
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Definition
| For proper pH & Mg, which is needed for polymerase |
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Term
| What is the temperature at which Maximum activity occurs with the Taq DNA polymerase? |
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Definition
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Term
| What is the temperature at which 10% activity occurs with the Taq DNA polymerase? |
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Definition
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Term
| What is the temperature at which activity gradually becomes inactivated with the Taq DNA polymerase? |
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Definition
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Term
| What is the optimum growth temperature for Taq DNA polymerase? |
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Definition
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Term
True or False:
Taq DNA Polymerase can survive higher temperatures than 70 degrees Celsius, including boiling water |
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Definition
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Term
| Taq DNA Polymerase is isolated from what? |
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Definition
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Term
| At what temperature does Denaturation occur? |
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Definition
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Term
| What happens during Denaturation? |
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Definition
| Separate strands by breaking hydrogen bonds |
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Term
| At what temperature does Annealing occur? |
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Definition
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Term
| What happens during Annealing? |
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Definition
Primer hybridization
Primers anneal to denatured DNA strands |
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Term
| At what temperature does Elongation occur? |
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Definition
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Term
| What happens during Elongation? |
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Definition
| Taq DNA Polymerase synthesizes new complementary DNA strands |
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Term
| How many times is the Cycle repeated? |
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Definition
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Term
| About how much time occurs per step? |
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Definition
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Term
| About how much time occurs per cycle? |
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Definition
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Term
|
Definition
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Term
|
Definition
| Heat (94 degrees Celsius) |
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Term
| What does Hydrogen bond formation require? |
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Definition
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Term
True or False:
The more G-C the lower the annealing temperature needs to be? |
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Definition
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Term
True or False:
The more G-C the higher the annealing temperature needs to be? |
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Definition
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Term
|
Definition
| Denaturation + Annealing + Elongation/Extension |
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Term
| With the Thermal Cycler how many cycles are performed in the same tube? |
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Definition
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Term
True or False:
The target sequence doubles with each cycle |
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Definition
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Term
| At the end of Cycle three how many fragments of desired length (precise length fragments) are present. |
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Definition
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Term
| At the end of Cycle four how many fragments of desired length (precise length fragments) are present. |
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Definition
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Term
True or False:
At the end of cycle 19 there have been over 500,000 DNA copies present. |
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Definition
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Term
| About how long does it take to make millions of copies of specific DNA sequence? |
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Definition
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Term
| What type of increase is seen in the number of target DNA sequences? |
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Definition
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Term
|
Definition
| PCR product used for detection and analysis |
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Term
| Standard PCR can amplify up to how many bp? |
|
Definition
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Term
| With modification, Standard PCR can amplify up to how many bp? |
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Definition
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Term
True or False:
Amplicons are abundant in PCR lab |
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Definition
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Term
| Preventing cross contamination is very important, otherwise what can happen? |
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Definition
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Term
What are the following considered:
Aerosols from popping top off microcentrifuge tube
Aerosols pulled into pipettes
Droplets on hand going everywhere |
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Definition
| Ways in which DNA spreads |
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Term
| What is in the Master Mix? |
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Definition
Taq DNA Polymerase
dNTPs
Primers
Buffer |
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Term
| What are the 3 work areas? |
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Definition
Reagent area
Extraction area
PCR area |
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Term
| What happens in the Extraction area? |
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Definition
| Patient samples are processed |
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Term
| Is work flow unidirectional or bidirectional? |
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Definition
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Term
| What is the direction of the work flow? |
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Definition
| Reagent --> Extraction --> PCR |
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Term
| You should store reagent in what volume? |
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Definition
| Small aliquots; discard aliquot after 1 use |
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|
Term
True or False:
Discard aliquot after 1 use |
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Definition
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Term
| What type of tips should you use for pipettes? |
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Definition
| ART (Aerosol Resistant Tips) |
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Term
| What kind of ware should you use to reduce contamination? |
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Definition
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Term
| What should you clean areas with? |
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Definition
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Term
| What will 2-10% bleach destroy? |
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Definition
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Term
| How many specimen tubes should you have open at a time? |
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Definition
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|
Term
True or False:
Keep tops on reagents as much as possible |
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Definition
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Term
| What should you do to the tubes before opening to decrease aerosols? |
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Definition
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