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.
Purifying Proteins
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 |
