Molecular Diagnostics 4 Flashcards
(38 cards)
Can you sequence genomic DNA directly?
no
what do you do first to sequece genomic DNA
isolate genomic DNA
PCR amplify
What do you use at template for sequencing DNA
PCR amplified DNA
What do you sequence for DNA
both alleles simultaneously
what is the issue with sequencing both alleles simultaneously for DNA sequencing
if there is mutation on one allele, whe you run a gell there will be two bands at the position there is a mutation
What is the mode of inheritance for beta thalassemia
AR
what regions does beta thalassemai have high incidence rate
mediterranean and africa (same as sickle cell)
Pt has hypochromia, osteoperosis abrnomalities, abnormalities of skull and bones, iron overload in liver, what is the disease
beta thalassemia
why is there iron overload in beta thalassemia
secondary to blood transfusions
what kind of mutation causes beta thalassaemia
null mutations
What kind of protein is produced with null mutation
no protein
If premmature stop codon is instroduced what will hapepn to protein
No protein
What is next generation sequencing
high-throughput, massively parallel. just a new technology that sequences very quickly, there are multiple kinds
What are the advanatges of next gen sequecing
very fast
less expensive
useful for whole genome resequencing
what are the disadvantages of next gen sequencing
not useful for de novo sequencing
high error rate
not targeted so can’t sequence a single gene
What techniques can be used to identify known mutations
RFLP
ASO
PCR
What techniques can be used to find changes in copy number (deletions, duplications)
FISH
CGH
MLPA
What techniques can be used to identify novel mutations
sequencing
If you don’t know anything about gene, where it is, where mutation is
allele-tracking
What is allele tracking
follow family pedigree and can follow how disease goes through pedigree
What are VNTRs
variable number of tandem repeats. also called short tandem repeats
what type of polymorphic marker is most used in allele tracking
CA-rpt marker, a type of VNTR
The higher the degree of polymophism, the what to eh population (about the marker)
the better the marker
How do you determine which of the 2 chromosomes is maternal or paternal in origin, etc. when doing allele tracking
use polymorphic markers
