Team:Chiba/Notebook/Calendar/18 September 2009



>Go to the Notebook page

(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

LasR is Unworking