MOLECULAR DIAGNOSTICS Flashcards
1
Q
Describe the steps that are involved in PCR reaction and their temperatures:
A
- denaturation- 95 degrees
- annealing- 50 to 60 degrees
- extension - 72 degrees
2
Q
What are the ingredients that are required in PCR:
A
- target DNA, primer, DNA polymerase, dNTP’s, machine to cycle temperatures- thermocycler, typically analyse products using gel electrophoresis
3
Q
What is an oligonucleotide?
A
- serves as primer for RNA synthesis
4
Q
What are the advantages of PCR?
A
- very specific assay determined by 2 primers
- start with very small amounts of DNA
- not dependant on DNA being intact
- can be used in combination with other techniques
5
Q
Describe quantitative PCR:
A
- based on detection and quantitation of a genetic reporter
- signal increases in direct proportion to the amount of product that is present
6
Q
Describe digital PCR:
A
- works by separating the mixture into an array of different partitions
- individual molecules are targeted and amplified
7
Q
What are the three applications of PCR:
A
- detection of infectious/genetic diseases
- Sex ID and screening of males versus females
- Genotyping
8
Q
Outline 3 applications of next generation sequencing:
A
- mixed breed identification
- understanding resistance to infection
- understanding disease transmission and vaccination requirements
- microbiome analysis
