Review of Basic Techniques and Purifying Proteins

Written by Kasia Bender

Objectives:

–       To practice calculating various solution components, including molarity and % (m/v).

–       To practice the use of a graduated pipet and micropipette.

–       To review lab safety.

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Part I A: Calculations

 

Part I B:

Table One:

Sample

Conc. (µg/µL)

glucose (µL)

buffer (µL)

1

0.0

0.0

187.5

2

1.0

30.0

187.5

3

2.0

60.0

187.5

4

3.0

90.0

187.5

5

4.0

120.0

187.5

6

5.0

150.0

187.5

Table Two:

Stock

MM vol.

MM conc.

Assay

Buffer

10 x

55 µL

1.25 x

1 x

DNTPs

2.5 mM

44 µL

0.25 mM

0.20 mM

Primer 1

62.0 µM

2.66 µL

0.375 µM

0.30µM

Primer 2

62.0 µM

2.66 µL

0.375 µM

0.30 µM

Water

335.68 µL

 

Part I C:

Table One: Graduated Pipet

Student 1

Student 2

Mass of microcenterfuge tube alone

1.05 g

1.02 g

Mass of microcenterfuge tube & water

2.05 g

2.03 g

Mass of water

1.00 g

1.01 g

 

Table Two: Micropipette

Student 1

Student 2

Mass of microcenterfuge tube alone

1.08 g

1.07 g

Mass of microcenterfuge tube & water

1.18 g

1.17 g

Mass of water

0.10 g

0.10