Genetics - The Future Of Medicine Flashcards
How is CF diagnosed?
Help prick test
electrophoresis after PCR = if affected one band which is lower
How to we diagnose CF because there are multiple mutations that can cause it?
Run PCR on whole gene and the pattern of fluorescence determines which mutation you have
How do we find out what the exact mutation is?
Sanger sequencing
Have a primer and add to DNA
Split it into 4 groups
Each reaction has a terminating nucleotide so no more extension
This means you will know what the end of each fragment is so you an put the sequence together
What is next generation sequencing?
Take DNA or RNA and fragment
Add on adapter sequences to end of fragment
Amplify this using primers that bind to adapters
Machine detects all the parts that are being amplified
How do we measure the tranlasome?
Ribosome profiling
Freeze all the ribsosomes
Digest rest of mRNA
And see where the ribosomes were sitting
How do we map the proteins produced by DNA?
Pull out RNA and do mass spec to see what the proteins are
OR
Pull out protein and see which RNA codes for it
Why do we map the genome?
Pharmocogenics for cancer treatment
Bio markers = diagnose by blood test
Identify cause if rare diseases
How is next generation sequencing used in diagnosing infectious diseases?
They have 16 s rRNA
This can be extracted and sequenced to see what it is
What is the minION?
RRNA passed through a nanopore and depending on what the base it changes the charge so it can read the sequence
What are the limitations of NGS?
Expensive
Library generation biases = some adapters bind easier than others
Read depth =the genes that are expressed a lot are not going to be too exciting but the ones that are rare we have to repeat a lot
Is the mutation detected even causing the disease?
What do bacteria do when they are infected by a bacteriophage?
Take up dna
Make hairpins and insert it into CRISPR region so when it is infected again it can chop up the new dna with teh one it already has
What is CRISPR?
Genome editing
Using their hairpins with any nucleotideGG we can change the genome
What are limitations of CRISPR?
NGG is quite common so too much changing
A lot of imperfect matches
