7.1.1 - PCR Flashcards
(15 cards)
What is a genome?
the total of all genetic material in an organism
What is a gene?
A section of DNA that codes for a protein
Who invented PCR and in when was the discovery made?
Kary B.Mullis
1983
What do you need for PCR?
Nucleotides, The DNA you want to amplify, Primer, Buffer solution, Taq polymerase
What is taq polymerase?
A type of DNA polymerase that is found in hot springs and is therefore stable at high temperatures
What are the 3 main steps in PCR?
Denaturation
Annealing
DNA synthesis
What happens in the denaturaton stage during PCR?
DNA is heated to 95 degrees, hydrogen bonds between chains break. 2 strands separate
What happens in the annealing during PCR?
Mixture is cooled to 50-65 degrees which allows primers to anneal/attach to 3’ end of each strand
What happens in the DNA synthesis stage of PCR?
Heated to 72 degrees for DNA polymerase to attach nucleotides. Heat tolerant taq polymerase replicates the region of DNA
What do primers do in PCR?
Allow polymerase to join and copy DNA strands
Binds to the section of DNA you want to amplify
What are two examples of when PCR is used?
To amplify copies of DNA found at a crime scene
Diagnosis of disease for example those caused by a virus
What is a real-time Reverse transcriptase PCR test?
Viruses have RNA as genetic material instead of DNA
Reverse transcriptase is used to convert RNA into DNA
A primer is added and the reverse transcriptase is used to convert RNA into cDNA
When would a real-time Reverse transcriptase PCR test be used?
To amplify the DNA of the virus that causes COVID
What common marking point comes up at the end of a 4/5 marker on PCR?
The cycle should be repeated several times
Describe the process of PCR?
Use of primers.
95 degrees used for 30 seconds to separate DNA strands.
55 degrees used for 20 seconds to allow primers to bind.
Heat to 72 degrees with taq polymerase to copy DNA.
Repeat process to obtain many copies of DNA.
