Restriction Enzyme Kinetics

Question 1

3 components of plasmid solution

Answer 1

6 ul 10 X cutsmart buffer
30 ul H2O
4 ul Xbal solution

Question 2

Agarose gel electrophoresis buffer preparation buffers

Answer 2

6X gel loading buffer
1X tris Acetate EDTA (TAE) running buffer

Question 3

2 components of gel loading buffer

Answer 3

0.0100g bromophenol blue
1.6g sucrose

Question 4

3 components of TAE running buffer

Answer 4

4.84g Tris
1.14g glacial acetic acid
2 ml of 0.5M EDTA ph 8.0

Question 5

Restriction enzyme used

Answer 5

Xbal

Question 6

Kinetics model

Answer 6

Michaelis-menten kinetics

Question 7

How does concentration of agarose affect DNA migration

Answer 7

Higher agarose concentration = smaller pores = slower migration

Question 8

Purpose of ethidium bromide (EtBr)

Answer 8

Added during separation of DNA fragments to visualize banding pattern/ stain DNA

Question 9

Charge lf EtBr

Answer 9

Positive charge = reduces DNA migration by 15%

Question 10

Why is dh5a e.coli used

Answer 10

High transformation efficiency

Question 11

Cut smart buffer composition

Answer 11

50 mM potassium acetate
20 mM tris-acetate
10 mM Magnesium acetate
100 ug/ml BSA ph 7.9
(Bovine serum albumin)