Practical Exam

Question 1

Instagene Matrix

Answer 1

removes metals/salts that inhibit PCR process

Question 2

Primers

Answer 2

Contain DNA fragments that bind during annealing step

Question 3

Taq polymerase

Answer 3

connects nucleotides together on a new DNA strand

Question 4

PCR buffer

Answer 4

contains salt and proper pH conditions for PCR

Question 5

nucleotides

Answer 5

free nucleotides that pair with complementary bases

Question 6

ice

Answer 6

keeps samples stable before PCR

Question 7

thermocycler

Answer 7

controls temperature changes for PCR

Question 8

gel box

Answer 8

used for running electrophoresis and compare PCR results

Question 9

E coli

Answer 9

white, rough, bigger

Question 10

S. aureus/staff

Answer 10

sunshine (yellow), smooth, small

Question 11

How to spread a plate properly

Answer 11

For a plate to be spread “properly” there must be isolated colonies and the quadrants should not run into each other.

Question 12

How do you know which tube has GFP

Answer 12

It glows when UV light is shined on it.

Question 13

Which would be more effective to purify a single protein from a mix of many proteins; affinity or HIC chromatography? Why?

Answer 13

Affinity chromatography because it uses ligands specific for a single target substance

Question 14

more colonies =

Answer 14

higher transformation efficiency

Question 15

lawn vs. colonies

Answer 15

glow vs. non-glow

Question 16

Why do we use blocking buffer?

Answer 16

After transferring the proteins from the agarose gel to the nitrocellulose membrane, if you do not add the blocking buffer, then the antibody can stick to all parts of the membrane, instead of just where the cow albumin is.