問題一覧
1
What is the first specific amplification method of any type was?
Polymerase chain reaction (PCR)
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What is the first and prototypical method for amplifying target nucleic acid?
Polymerase chain reaction (PCR)
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Who developed PCR?
kary mullis mid-1980’s
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This allows the rapid synthesis of designated fragments of DNA
PCR
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The main goal of PCR is to create multiple copies/ amplify target (DNA/RNA) sequence
true
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AKA “copy machine” for DNA that revolutionized molecular biology
PCR
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Target amplification involves making MANY COPIES of a specific DNA sequence
amplicons
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PCR is DNA replication in vitro
true
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What is the main replication enzyme of PCR?
DNA polymerase
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What are the components of PCR?
water, buffer, DNA template, Thermostable polymerase, primer pair, magnesium, deoxynucleotides triphosphates (dNTPs)
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Components of PCR is also known as?
AOTA
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Stabilizes the DNA polymerase, DNA and nucleotides
buffer
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Containes the target region of interest
DNA template
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What is the critical componen of PCR?
primer pair
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Primer pair is also known as?
Oligodeoxynucleotides
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This stabilized the DNA double helix in components of PCR
magnesium
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Equimolar building blocks of DNA
dNTPs
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Preparation of PCR reaction components (mastermix)
Pre PCR
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Isolation of nucleic acid from samples
Pre PCR
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Thermal cycling (denaturation, annealing, extension)
PCR
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Analysis of amplified DNA
post PCR
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What is seen in the picture?
typical PCR set-up in laboratory
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High heat to disrupt and separated DNA duplexes
Heat denaturation (PCR)
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Attachment of primers to the template strand
primer annealing (PCR)
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Elongation of daughter strand with DNA polymerase
primer extension (PCR)
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This control in PCR ensures that the ENZYME IS ACTIVE , the buffer is optimal, the primers are priming the right sequences, and the thermal cycler is cyling appropriately
positive control
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Aka contamination control or reagent blank which ensures that the reaction MIX IS NOT CONTAMINATED WITH TEMPLATE DNA or amplified products from a previos run
negative control without DNA
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Lacks the target sequence, ensures that the primers are NOT ANNEALING to nontarget sequences of DNA
negative control with DNA
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RT stands for?
reverse transcription
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rt stands for?
real time
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This refers to the CONVERSION OF RNA TEMPLATE into its complementary DNA strand (cDNA)
Reverse transcriptase PCR
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This is a POWERFUL TECHNIQUE that enables amplification of TWO or MORE products PARALLEL in a single reaction tube.
Multiplex PCR
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What do you call this?
cold rack
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What do you call this?
PCR strips
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What do you call this?
PCR plates
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What do you call this?
centrifuge