DLA 17 + lecture 18 Flashcards
How can enzymes be regulated?
- concentration of the substrate/product
- modulation of enzyme concentration
- covalent modification of enzymes
- allosteric regulation
What are the different kinds of hybridization methods?
- sequencing (primer)
- Southern blot
- Northern blot
- RFLP analysis
- ASO probes
- Western blots
When DNA is tested, what is the main purpose?
Look for mutations in the coding sequence
Why use a southern blot?
A southern blot is used to look for deletions and insertions in the DNA
Why use a northern blot?
The sample only contains mRNA molecules that have been isolated from the cell
What is a western blot used for?
Detect and quantify the amount of a particular protein
What are the steps to the di-deoxy method?
- add DNA to be sequenced
- denature the DNA to expose the template
- add radioactively labelled primer
- add dNTPs
- Make 4 aliqouts
- add dideoxy NTPs
- add enzymes
What is RFLP analysis?
Restriction Fragment Length Polymorphism
Used to detect base pair changes that impact restriction sites
What are the two ways to do RFLP?
It used to be done by the southern blot but now it is done by PCR
ASO probes?
they are normally 15-21 base pairs long
used to detect polymorphisms or common genetic mutations
CFTR and sickle cell
What test to use if the sample size is small?
amplify
What test to use if it is a common mutation?
ASO or RFLP
What test to use if the sample has a restriction site change
RFLP
What test is it when the example shows a spot
ASO
what test to use if it is a known family mutation?
RFLP
