Additional Questions Flashcards
1
Q
Restriction Endonucleases
A
- a.k.a. restriction enzymes
- Methalated sequences on host bacteria DNA prevent cutting of host DNA
2
Q
What is TBE Buffer?
A
- This is the buffer, fills the pores in the arose gel, so that DNA fragments can diffuse through
- You add this before you place your samples into the wells
3
Q
What is Loading dye? (what is it made of and what does it do?)
A
- Glycerol: to make the sample sink to the bottom of the well
- Bromophenol Blue: this enables us to see the sample, and tell when it is reaching the end of the arose gel, so we know when to stop electrophoresis
4
Q
Cutting sequence of Alu 1
A
AG CT
TC GA
5
Q
Cutting sequence of Taq 1
A
T CGA
AGC T
6
Q
Cutting sequence of EcoR1
A
G AATTC
CTTAA G
7
Q
Cutting sequence of Not 1
A
GC GGCCGC
CGCCGG CG
8
Q
Ethidium Bromide: what does it do, what colour is it?
A
- Used to visualise the DNA
- when bound between bases, it takes on a pink-orange fluoresce under UV light
9
Q
Transaminases: cofactor? what do they do?
A
- Pyridoxial phosphate: from Vitamin 6
- Move amino groups from amino acid and oxo acids
10
Q
What was the transaminase reaction we did? And with what enzyme?
A
- Glutamate (A.A.) + Pyruvate (keto.a.) –> Alanine (A.A.) + Oxoglutarate (keto.a.)
- Glutamate amino transferase
11
Q
Glutamate Dehydrogenase: cofactor? what does it do?
A
- Catalyses oxidative deamination of glutamate into oxoglutarate
- NAD+ and NADP+ as a cofactor
12
Q
What does aminopeptidase do?
A
- Cleaves the amino acid off the N terminus
13
Q
What does carboxypeptidase do?
A
- cleaves the amino acid off the C terminus
14
Q
2,4,-DNP
A
- Stains keto/oxo acids yellow
15
Q
Ninhydrin + Heat
A
- Stains Amino Acids purple
