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Started on Monday, 14 October 2019,

12:51 AM
State Finished
Completed on Monday, 14 October 2019,
12:55 AM
Time taken 3 mins 49 secs
Marks 12.00/12.00
Grade 100.00 out of 100.00

Question 1
Correct

Mark 6.00 out of 6.00

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This week you will isolate plasmid DNA from two

white colonies. The question below lists all the
di!erent steps in the procedure. Put these into
the correct order for successful isolation of
plasmid DNA.

6
Add bu!er 300 µl of Bu!er A3.
!
Pipette supernatant into top part 9
of Spin Column Assembly.
!
Add 50μL of Elution Bu!er to
Spin Column Assembly and 14
centrifuge (≥11,000 x g) for 1
minute. !
Decant and make sure pellet is 2
as dry as possible.
!
Remove and discard top part of 15
Spin Column Assembly.
!
Mix gently with bu!er containing
4
SDS and sodium hydroxide to
lyse the cells. !
Centrifuge precipitated cell
8
debris and chromosomal DNA
for 5 min (≥11,000 x g). !
Centrifuge Spin Column 10
Assembly for 1 min (≥11,000 x g).
!
Incubated at room temperature 5
for 5 minutes.
!
Re-assemble Spin Column
Assembly and dry silica 12
membrane by centrifugation (1
minute, ≥11,000 x g).
!
Spin bacterial suspension for 30 1
s in microcentrifuge.
!
Mix by inverting the tube 7
multiple (~10) times.
!
Remove Spin Column from
11
centrifuge, decant waste solution
and add 600μL wash bu!er A4. !
Resuspend cell pellet in 250μL 3
Bu!er A1.
!
Place top part of Spin Column 13
Assembly onto Collection Tube.
!

Question 2
Correct

Mark 4.00 out of 4.00

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The following solutions are provided:

10x restriction enzyme bu!er,
PstI (5 units/µL),
pUC19 DNA 10mg/mL and
Sterile water.
Calculate the amounts of each of the components
you would need to digest 10µg of pUC19 DNA in a
total volume of 50µL. Note: Use 5 units of PstI in a
total of 50µL.

10X Restriction enzyme 5 µL

bu!er
!
1 µL
pUC19 DNA
!
43 µL
Sterile water
!
1 µL
PstI
!

Question 3
Correct

Mark 2.00 out of 2.00

Flag question

If 31 µL of pUC19 DNA contain 14.91 µg of DNA.

What is the concentration (in µg/µL) of pUC19 in
this solution? Give your answer to the nearest 2
decimal places.

Answer:

0.48ug/uL !

Finish review

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