Experiment Overview/Pipetting

Question 1

types of micropipettes?

Answer 1

1. gilson (2 stops)
2. eppendorf (3 stops)

Question 1

what are the steps of the experiment?

Answer 1

1. Restriction digest of insert (LDL receptor)
2. Transformation into E. coli JM101
3. determine cholesterol conc in spent media via standard curve
4. RD and plasmid map

Question 2

1st thing to do when setting pipette?

Answer 2

check range

Question 3

range and reading of each type of micropipette?

Answer 3

p2: 0.2µL-2µL (1 black digit, 2 red decimal places)
p20: 2-20µL (2 black digit, 1 red decimal place)
p200: 20-200µL (all black no decimal place)
p1000: 200/100-1000µL (highest is 100 = 1000, 20 = 200)

Question 4

if 2nd stop is used to draw liquid..

Answer 4

too much volume is drawn

Question 5

using pipette out of range causes…

Answer 5

1. inaccurate volume measurements
2. damage to internal pipette mechanisms

Question 6

using pipette without tip causes…

Answer 6

1. contamination
2. pipette corrosion

Question 7

What are the pipette tip types?

Answer 7

• yellow - P20 + P200
• blue - P1000
• filter tip - contains sterile filter within each tip to preent contamination