User:Karmella Haynes/Notebook/ASU courses/2012/01/30

From OpenWetWare

Jump to: navigation, search
ASU Courses Main project page
Previous entry      Next entry

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 =

  1. Cuvette 1: O2-GFP (positive control + 4.0 mM IPTG)
  2. Cuvette 2: O#-GFP/ LacI (0.0 IPTG)
  3. Cuvette 3: O#-GFP/ LacI (0.25 IPTG)
  4. Cuvette 4: O#-GFP/ LacI (0.5 IPTG)
  5. Cuvette 5: O#-GFP/ LacI (1.0 IPTG)
  6. Cuvette 6: O#-GFP/ LacI (2.0 IPTG)
  7. Cuvette 7: O#-GFP/ LacI (4.0 IPTG)
  8. 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

  1. Banana, no IAA
  2. Banana, + 5 uL IAA


Personal tools