Week 7 Tuesday

Question 1

What would a mutation look like?

Answer 1

The sample would have a base pair that is not correct (e.g G paired with A)

Question 2

What would a deletion look like?

Answer 2

The sample would have a - where a base would be

Question 3

What would an insertion look like?

Answer 3

The sample has extra bases and the reference has —

Question 4

What is the reference sequence?

Answer 4

what you think the sequence is (the hoped result)

Question 5

If we couldnt perform DNA sequencing, what other methods could we use?

Answer 5

plasmid size- run on a gel
Restriction Digest map
PCR confirmation- amplify a piece that is only on one plasmid
Assess activity (de genome editing and if GFP is in there, you wont be able to)

Question 6

What are the pros and cons of running a plasmid on a gel?

Answer 6

pros: quick and easy
cons: misses mutations. If we were replacing GFP with a sequence that was the same size as GFP, it wouldn’t be able to differentiate or would be hard to read

Question 7

What are the pros and cons of a restriction digest map?

Answer 7

pros: distinguished plasmids of similar size
cons: misses mutations, requires planning, time, and using up plasmid

Question 8

What are the pros and cons of PCR confirmation?

Answer 8

pros: distinguished plasmids of similar size
cons: misses mutations, planning, time, and have to order a primer

Question 9

What are the pros and cons of just assessing activity?

Answer 9

Pros: saves time if it works, dont have to add a control to test for
cons: wastes time and money if it doesn’t work

Question 10

What does assessing the activity mean?

Answer 10

transforming the plasmids into the yeast and seeing what happens