Activity 2.1.2

Question 1

What is DNA extraction?

Answer 1

The process of isolating and purifying DNA

Question 2

What does DNA extraction entail?

Answer 2

Removal of all cell structures and enzymes

Question 3

What does potassium acetate do?

Answer 3

Precipitates non-DNA material

Question 4

What does precipitate mean?

Answer 4

Chemical process that causes a substance to deposit in a solid form

Question 5

What is a supernatant?`

Answer 5

The liquid lying above a solid residue after crystallization, precipitation, centrifugation, or other process

Question 6

What is a pellet?

Answer 6

The clump of materials that forms at the bottom of the micro-centrifuge tube after centrifugation

Question 7

What is the first goal of the lab?

Answer 7

Centrifugation

Question 8

What is the second goal of the lab?

Answer 8

Removal of supernatant

Question 9

What is the third goal of the lab?

Answer 9

Addition of lysis buffer, mixing/incubation

Question 10

What does the lysis buffer contain?

Answer 10

Detergents to disrupt cell membrane

Question 11

What is the fourth goal of the lab?

Answer 11

Addition of potassium acetate

Question 12

What is the fifth goal of the lab?

Answer 12

Addition of isopropanol

Question 13

What does isopropanol do?

Answer 13

Precipitates DNA, removal of salts

Question 14

What is the sixth goal of the lab?

Answer 14

Removal of supernatant from pellet

Question 15

What is the seventh goal of the lab?

Answer 15

Addition of 70% ethanol

Question 16

What is the last part of the lab?

Answer 16

Remove supernatant, allow pellet to dry, add Tris buffer

Question 17

What does the Tris buffer do?

Answer 17

Elutes/dissolves DNA –> DNA back in solution

Question 18

What is the ultimate product of the lab?

Answer 18

Purified DNA

Question 19

How is a multiplex PCR different than a standard PCR?

Answer 19

It uses multiple primers in order to produce amplifications specific to different DNA sequences, allows you to test for multiple bacteria at once

Question 20

Is this test a preliminary test or conclusive test?

Answer 20

Conclusive because it identifies a specific strain of bacteria instead of eliminating possible types

Question 21

If you were to run a multiplex PCR and saw three bands (in the electrophoresis gel) from a sample suspected to be contaminated with pathogenic bacteria, what can you conclude about the sample?

Answer 21

You could conclude that the sample was, in fact, contaminated by three different strands of bacteria