DNA EXTRACTION

Question 1

Acquire the bacterial stock culture, gram negative bacteria

Answer 1

1

Question 2

Transfer 1mL of specimen to 1.5 mL microcentrifuge tube

Answer 2

2

Question 3

Centrifuge tube at maximum speed for 1 min

Answer 3

3

Question 4

Discard supernatant

Answer 4

4

Question 5

Add 100 uL of EL buffer then pipette mix

Answer 5

5

Question 6

Incubate at 37’C

Answer 6

6

Question 7

Add 100 uL of RS buffer and pipette mix

Answer 7

7

Question 8

Add uL of PK and pipette mix

Answer 8

8

Question 9

Incubate at 56’C for 15 minutes

Answer 9

9

Question 10

Add 200 uL of GA buffer

Answer 10

10

Question 11

Centrifuge at 13,000 KG for 1 minute

Answer 11

11 and 15

Question 12

Transfer supernatant to new 1.5 mL microcentrifuge tube

Answer 12

12

Question 13

Add 400 uL of BA buffer and pipette mix

Answer 13

13

Question 14

Transfer mixture to spin column

Answer 14

14

Question 15

Add 500 uL of G Binding buffer

Answer 15

16

Question 16

Cetrifuge at 10 000 KG for 1 minute

Answer 16

17, 20, 23, 27

Question 17

Discard flowthrough

Answer 17

18, 21

Question 18

Add 500 uL of wash buffer

Answer 18

19

Question 19

Transfer spin column to a new 1.5 microcentrifuge tube

Answer 19

24

Question 20

Add 100 uL of elution buffer

Answer 20

25

Question 21

Incubate at RT for 1 minute

Answer 21

26

Question 22

The purified DNA extract is now in the receiving vessel

Answer 22

28