01/30/12
- (BME 194) Section A/ Section B classes
- (BME 194) Experimental cultures for Section C
Class prep (1 hour ahead)
> 1 set of cuvettes =
- Cuvette 1: O2-GFP (positive control + 4.0 mM IPTG)
- Cuvette 2: O#-GFP/ LacI (0.0 IPTG)
- Cuvette 3: O#-GFP/ LacI (0.25 IPTG)
- Cuvette 4: O#-GFP/ LacI (0.5 IPTG)
- Cuvette 5: O#-GFP/ LacI (1.0 IPTG)
- Cuvette 6: O#-GFP/ LacI (2.0 IPTG)
- Cuvette 7: O#-GFP/ LacI (4.0 IPTG)
- Cuvette 8: LacI (4.0 IPTG)
Exercise 1: GFP measurement: Vikram's spectral analyzer; fill 2 mL cuvettes w/ 1 mL culture
- Section A (Monday morning, 9 am)
- Switch A (O2-GFP) - 2 sets
- Switch B (O1-GFP) - 2 sets
- Switch C (Osym-GFP) - 1 set
- Section B (Monday afternoon, 3 pm)
- Switch A (O2-GFP) - 3 sets
- Switch B (O1-GFP) - 2 set
- Switch C (Osym-GFP) - 3 set
Students record data on worksheet #1
Exercise 2: Students also measure OD600 on spectrophotometer
Exercise 3: Students take pictures of streaked agar plates using Vikram's web cam with filters
Experimental cultures for Tuesday section
> Innoculate overnight cultures w/ 10 uL starter culture (20 tubes, 10 mL each)
|
|
Medium
|
IPTG (mM)
|
| O2-GFP |
Amp |
|
|
|
|
|
4.0 (2x)
|
| O2-GFP/ LacI |
Amp/ Kan |
0.0 |
0.25 |
0.5 |
1.0 |
2.0 |
4.0
|
| O1-GFP/ LacI |
Amp/ Kan |
0.0 |
0.25 |
0.5 |
1.0 |
2.0 |
4.0
|
| Osym-GFP/ LacI |
Amp/ Kan |
0.0 |
0.25 |
0.5 |
1.0 |
2.0 |
4.0
|
| LacI |
Kan |
|
|
|
|
|
4.0 (2x)
|
Banana induction: 5 mL each
- Banana, no IAA
- Banana, + 5 uL IAA
| 
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