Lab techniques Flashcards
(17 cards)
What examples of standard molecular techniques?
Karyotyping
Blotting
Sangar sequencing
PCR
What does karyotyping allow to find?
Translocation
application and deletion
ploidy
What phase of the cell in karyotyping done?
M phase
What does FISH allow that Karyotyping doesn’t?
more sensitive
probes can find specific DNA sequences
Can use interface chromosomes
What is Comparative genomic hybridisation?
Type of karyotyping. Raddiolables for normal and tumour DNA. shows where the present of tumour sequences on chromosomes or in an array.
What is blotting?
taking of DNA/RNA/ protein and using electrophoresis
What type of blotting uses protein?
Western
What does southern blotting?
DNA analysis, can show presence of specific gene.
What does Northern blotting?
RNA analysis, shows gene expression and protein size
What are the stages of PCR?
- Denaturation
- Annealing
- Extension
- Completion
What can PCR be used for?
Amplification
Presence of minimal residual disease
What examples of advanced techniques are there?
Qualitative PCR
droplet digitalPCR
Microarray
next generation sequencing
What is the advantage of quantitive PCR?
Real time quantification of DNA with every cycle
How can a PCR be quantitive?
Florescent probes show conc of DNA. Exponential phase of DNA product increase is proportion to the initial input DNA.
What disease is quatitivative PCR use full in?
BCR-ABL (Philadelphia chromosome) can be used to monitor leakaemia
Explain how droplet digital PCR
First PCR
then droplets with fourescent probes attached
Flourescence measured
How is a DNA microarray performed?
- Many DNA probes on array
- Hybridisation of array with sample DNA
- array read for probes
