Genomic Medicine Flashcards
Whole genome sequencing
Looking at all (3billion) DNA base pairs
Whole exome sequencing
Only look at the sequence of the coding genes.
RFLP analysis
Restriction fragment length polymorphism- looking at one specific region of DNA
Uses restriction enzymes that cleave DNA molecule at unique sites depending upon sequence.
Can be used to diagnose
Ex. sickle cell anemia– if treat DNA with enzyme it will cut the two strands into different fragments.
STRP analysis
Short tandem repeat analysis. Used to determine different stretches of DNA (not whole thing)
Can be used as identifier- everyone has unique numbers of sequences.
SNPs
Single nucleotide polymorphisms
look at single nucleotide changes in a population
Used to compare diseased DNA to normal healthy DNA
Agrose gel electrophoresis
DNA inserted at top (negative/cathode) then electric current applied.
Longer pieces will remain closer to top. Smaller mover further
Southern Blot
Take gel membrane and soak with a specific DNA probe
Use x ray film to try to determine if a specific stretch of DNA is present
PCR
Amplifies a segment of DNA
Use a template DNA to bind to the oliggonucleotide primers.
Design primers to recognize primers.
Separates strands and uses DNA polymerase.
Used for detecting infections, cancer screening, genetic screens. Tissue matching.
Northern Blot
Used to detect RNA.
Run on gel, transfer to membrane and use probes to detect.
Microarray
Chip with sequences on it that RNA will bind to.
When they bind on each site it will glow. Computer program tells what gene it is.
Can look at which genes are being expressed and compare healthy to cancer.
Maybe try to make tailored treatment and predict what will work.
RNA seq
See the expression and sequence of the transcripts.
Like microarray but with sequencing also.
Gel Based Proteomic analysis
Add proteins to the gel, denaturing proteins and running out proteins that will separate out according to molecular weight.
Used to see protein in the cell because the gene might not express it even if it is there.
Gel Free proteomic analysis methods
MASS SPEC
Take an unknown sample and utilize to determine what the chemical nature of the protein is.
CRISPR gene editing
Machinery in bacteria that chew up certain bacteriophage at specific sites.
We exploit that machinery and tailor it to target and edit certain gene sequences.
CAS9- cuts double strand DNA
single guided RNA brings CAS9
Uses template to fix the break.
What gene was targeted in the embryo pathogenic gene correction?
MYBPC3, people end up developing hypertrophic cardiomyopathy.
Injected zygote with CRSPR
CAS 9- nuclease that induces double strand breaks.
The wild type copy of gene in early embryo was used as template
