MMBIO 241 Test

Question 1

What are the four factors that affect enzyme velocity

Answer 1

temperature, enzyme concentration, substrate concentration

Question 2

Explain the procedure of isolating acid phosphatase.

Answer 2

extract from wheat germ. Smash up wheat germ in extraction buffer, centrifuge, take supernatant which contains the enzyme.

Question 3

When you want to determin Vmax and Km?

Answer 3

vary substrate concentration, hold time and enzyme concentration constant. graph substrate concentration versus enzyme velocity

Question 4

to get an accurate value for V0

Answer 4

vary time and hold substrate concentration and enzyme concentration constant. Graph time vs. product concentration

Question 5

What is happening at Vmax?

Answer 5

enzyme is saturated

Question 6

what does km represent?

Answer 6

high dm means low affinity. low km means high affinity

Question 7

whats the purpose of pcr?

Answer 7

amplify a DNA target sequence

Question 8

What are the three steps in PCR?

Answer 8

denaturation, primer annealing, elongation

Question 9

Who long is PV92?

Answer 9

300 bp long

Question 10

What reagents are needed in PCR?

Answer 10

instagene matrix (chelating agent; prevents DNAse from degrading – Protease K), primers, Taq polymerase, PCR buffer, nucleotides

Question 11

How do you keep your sample from degrading before you run PCR?

Answer 11

ICE ICE BABY

Question 12

How is Pure Culture used?

Answer 12

single ancestor…. spreading?

Question 13

How is selective media used?

Answer 13

test against antibiotic

Question 14

GFP is only activated int he presence of?

Answer 14

Arabinose

Question 15

What is the formula for transformation efficeincy?

Answer 15

(# colonies)*(dilution factor)/ug plasmid (or DNA)

Question 16

How do you find dilution factor?

Answer 16

total volume of mixture/volume you spread on plate

Question 17

Describe how to do GFP chromotography

Answer 17

bacteria suspended in buffer, add lysozyme, incubate on dtry ice, centrifuge, keep supernatent, run supernatent in hydrophobic chromotgraphy

Question 18

Just the chromotography part

Answer 18

Equilibriation buffer, GFP sample, wash buffer, elution buffer

Question 19

Steps in our western blot?

Answer 19

run sera from different organisms on native agarose gel, transfer proteins to nitrocellulose membrane, block with gelatin, add antibodies, add color devlopment soultion that causes the enzyme on the antibody to color puprple

Question 20

darker bands on western blot mean what?

Answer 20

more closely related

Question 21

how do you know what organisms are more closely related in a phylogenttic tree?

Answer 21

closer together. branch length on the phylogenetic tree does count