lecture 4 Flashcards
what does genotyping an individual determine?
which alleles an individual carries for a specific gene (or set of genes)
what does genotyping of pathogen allow?
diagnosis of species and strain of infecting pathogen
what does phenotyping the disease measure?
‘taking a snapshot in time’, measures disease progression + severity
when genotyping an individual, what are sources of DNA?
blood, hair, buccal smear, cells from aminiotic fluid
which 2 PCR-based techniques are used for genotyping an individual?
PCR-RFLP - restriction fragment polymorphism
ARMS-PCR - amplification refractory mutation
what is the PCR-RFLP process?
- amplify the substrate (dsDNA) - then has a restriction enzyme site to cleave DNA
- add the restriction enzyme
what happens if RE site isn’t present in dsDNA?
no break
what happens if RE site is present in one dsDNA?
one will break
what happens if RE site is present in both dsDNA?
both will break
what is an example of PCR-RFLP?
diagnosis of Sorsby’s fundus dystrophy
what is Sorsby’s fundus dystrophy?
mutation in TIMP3 - introduces stop codon
degenerative eye disease leading to blindness (autosomal dominant)
what is ARMS-PCR function?
-detects any mutations involving single base changes
what is made in ARMS-PCR?
2 primers - 1 for each allele - binds specifically
what are the steps in ARMS-PCR?
- primer design
- amplification
- electrophoresis
- results
what can ARMS-PCR detect?
cystic fibrosis
