lab4

Question 1

Site-directed mutagenesis is a method by which

Answer 1

specific changes can be introduced into known DNA sequences

Question 2

Site-directed mutagenesis is used to study _________

Answer 2

the effects of particular bases on DNA structure or the ability of a DNA sequene to act as a transcriptional control element.

Question 3

The Q5 Site-Directed Mutagenesis method uses

and thus allows mutagenesis to be carried out ______

Answer 3

double-stranded DNA

directly in a plasmid

Question 4

the Q5 Site-Directed Mutagenesis method requires a pair of _____________
and these _____ to the template and are ___________ by means of ________

Answer 4

complementary mutagenic primers that contain a mismatch with the template
anneal
extended during temperature cycling
Q5 Hot Start High Fidelity DNA Polymerase

Question 5

Dpn1 is specific for ___________
So the _________, isolated from E. coli is ________ and subject to ________
The digested parental plasmid dna is not _____
and so the _______ _______ ________ ______ DNA is then transformed preferentially into the cell strain

Answer 5

methylated and hemimethylated dna
original plasmid DNA
methylated
digestion
transformed

Question 6

GFP’s values

Answer 6

ability to fluoresce in foreign organisms in the absence of substrates and A-victoria specific enzymes

Question 7

some of the more popular applications of GFP are

Answer 7

as a reporter gene, and as a fusion protein to study protein function and subcellular localization