POLYMERASE CHAIN REACTION

Question 1

An ______ method for amplifying DNA segments that depends on repeated cycles of denaturation, primer annealing, and DNA polymerase-directed DNA synthesis

Answer 1

In vitro - Polymerase Chain Rxn

Question 2

Rapid creation of a large quantity of product (_____) from very small quantities of the DNA template

Answer 2

amplicons - PCR

Question 3

PCR is developed by ____ in _____

Answer 3

Karry Mullis - 1983

Question 4

PCR is discovered while working through a ________

Answer 4

mutation-detection method

Question 5

First successful amplification (PCR)

Answer 5

Escherichia coli plasmid, pBR322

Question 6

First paper of practical application (PCR)

Answer 6

amplification of beta-globulin and analysis for diagnosis of sickle cell anemia

Question 7

PCR is called the method “________________” because _________ was the enzyme used to drive the replicationof DNA in a chain rxn

Answer 7

Polymerase catalyzed chain reaction - DNA polymerase

Question 8

PCR is classified under a larger group of molecular techniques known as “____________”

Answer 8

Nucleic Acid Amplification Techniques (NATs)

Question 9

Types of NATs

Answer 9

TPS

1. Target
2. Probe
3. Signal

Question 10

Tests under TARGET AMPLIFICATION

Answer 10

PTG

PCR, Transcription-based amplification system, Genome amplification methods)

Question 11

Tests under Probe Amplification

Answer 11

LQS

Ligase chain rxn, Strand displacement amplification, QB(eta) replicase

Question 12

Tests under Signal Amplification

Answer 12

BHCC

Branched DNA amplification, Hybrid capture assays, Cleavage-based amplification, Cycling probe

Question 13

Involves making many copies of specific DNA sequence

Answer 13

Target Amplification

Question 14

this method of cloning DNA using vectors and host cells is LABOR INTENSIVE and TIME-CONSUMING

Waiting for cells to replicate at detectable levels, it can take __________, whereas replicating the nucleic acid in vitro only takes ________

Answer 14

days to weeks or months - minutes to hours - Target amplification

Question 15

Application of PCR

Answer 15

• Screen mutations
• Detect bacteria and viruses in humans and pathogenic bacteria in contaminated food
• Collect DNA from extinct species
• Comparison of DNA
• Forensics (paternity testing, crime scene, ID)
• Production of DNA segments

Question 16

Principle of PCR

Based on the principles of DNA replication IN VIVO- AUTOMATED VERSION of DNA replication, performed in a test tube containing a total reaction volume of _______

Answer 16

20-50 mL