Team:Chiba/Notebook/Calendar/18 September 2009

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(17_September_2009 <|>19_September_2009)

LuxR Mutants Sequence

Purification of DNA

  • isopropanol precipitation and analysis
  1. add isopropanol(75%) 50 uL to PCR solution made yesterday
  2. VORTEX and centrifugation(14000 rpm 10 min)
  3. put off the supernatant fluid
  4. add isopropanol(75%) 50 uL
  5. centrifugation(14000 rpm 10 min)
  6. put off the supernatant fluid
  7. drying 60 degrees C 15min
  8. add ddH2O 20 uL and VORTEX
  9. sequence analysis

Las Check

Culture Mix

  1. K084007:LasI
  2. K084012:LuxI
  3. T9002:LuxR-GFP
  4. K091134:LasR-GFP
  5. I731012:LuxR-RFP


Sender:Receiver=0.8 mL:0.8 mL

(1):1 + 4 ---LasI/LasR-GFP

(2):1 + 3 ---LasI/LuxR-GFP

(3):1 + 5 ---LasI/LuxR-RFP


(4):2 + 4 ---LuxI/LasI-GFP

(5):2 + 3 ---LuxI/LuxR-GFP

(6):2 + 5 ---LuxI/LuxR-RFP


(7):4 + flesh LB ---LasR-GFP

(8):3 + flesh LB ---LuxR-GFP

(9):5 + flesh LB ---LuxR-RFP



Result

sample fluorescence
(1) -
(2) +
(3) +
(4) -
(5) ++
(6) +
(7) -
(8) -
(9) +


LasR is Unworking

Lab Notebook

Chiba-Labwork-17Sep09.jpg Chiba-Labwork-18Sep09.jpg Chiba-Labwork-17Sep09-5.jpg Chiba-Labwork-17Sep09-2.jpg