Lab 23

Question 1

What do both yeast variants require

Answer 1

leucine, uracil, histidine, tryptophan, lysine

Question 2

Focus lab 23

Answer 2

verify by PCR that partial deletion of RFP gene was made in RJY29 chromosome

Question 3

Where/what is the PAM sequence

Answer 3

protospacer adjacent motif on target DNA tells Cas9 where to cut (3 upstream) - NGG sequence

Question 4

DNA primers are always written in the ________ direction

Answer 4

5’-3’

Question 5

What does KCl do in the master mix?

Answer 5

stabilize primer - template binding

Question 6

MgCl2

Answer 6

cofactor for enzyme, neutralizes DNA charge

Question 7

dNTPs

Answer 7

substrate for enzyme

Question 8

5% Glycerol

Answer 8

stabilizes DNA pol and lowers temp required for denaturation

Question 9

IGEPAL CA-630

Answer 9

Non-ionic detergent stabilizes DNA polymerase

Question 10

Tween 20

Answer 10

Non-ionic detergent stabilizies DNA pol and prevents inhibition by ionic detergents

Question 11

Orange G and Xylene Cyanol

Answer 11

Tracking dyes

Question 12

Taq DNA Pol

Answer 12

Enzyme for DNA synthesis can withstand denaturing heat