PCR Flashcards
What does PCR stand for?
Polymerase chain reaction
Who invented PCR and when?
Kary Mullis 1985
How many bases are oligonucleotides?
18 bases
What 5 things are needed
Temple DNA DNA polymerase Oligonucleotide primer dNTPs Buffer
Where do primers bind to DNA?
At the 5’ end, the primer binds
What are amplicons?
Amplified DNA
What is the name of the DNA products?
dsDNA products
Name the 3 steps in PCR
1) Denature template DNA to ssDNA heat
2) Lower Temp to allow Annealing of primers
3) Extension of the primer by DNA polymerase
What are the temperatures of the steps in PCR?
95
55
72
Each cycle what happens to the DNA
dsDNA doubles
What bacteria is used and why?
Thermus Aquaticus
optimum temp of 72C
Relatively stable at 94C
How can we tell gene changes in gel electrophoresis?
Further up or down the page indicates deletion or insertion
What information is gained from a Prenatal test?
Detects mutated gene
What information is gained from a Prenatal screen?
Embryo tested across population for an increased risk of condition
What information is gained from a new-born screen?
Population wide testing for several treatable inborn errors of metabolism
