Team:Wageningen UR/Notebook/Proj1/July

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(Difference between revisions)
(→July - Synchronized Oscillatory System)
(→July - Synchronized Oscillatory System)
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RBS-GFP
RBS-GFP
-
Cultures were put in a shake incubator at 30°C, 300 rpm.
+
Cultures were put in a shake incubator at 30°C, 300 rpm. Temperature was set to 30 degrees as GFP is degraded at higher temperatures.  
The OD600s of every culture were measured 2 hours after inoculation and subsequently each hour, by taking 1 mL samples from the culture. the OD was measured with a Hitachi U1500 spectrophotometer. LB+amp was taken as blank.  
The OD600s of every culture were measured 2 hours after inoculation and subsequently each hour, by taking 1 mL samples from the culture. the OD was measured with a Hitachi U1500 spectrophotometer. LB+amp was taken as blank.  
Fluorescence was measured in a 96-wells plate on a fluorescent plate reader (Molecular Devices, M2 (toxicology). First blank measurements were done with 96 wells plates containing 50 ul LB+amp. The output corresponding with the specific wells was saved and used for further calculations.  
Fluorescence was measured in a 96-wells plate on a fluorescent plate reader (Molecular Devices, M2 (toxicology). First blank measurements were done with 96 wells plates containing 50 ul LB+amp. The output corresponding with the specific wells was saved and used for further calculations.  
 +
OD600 data presented below
 +
 +
{|! border="1" align="center" style="text-align:center;"
 +
! Sample
 +
! no. 1 (2 hours)
 +
! no. 2 (2 hours)
 +
! no. 1 (4 hours)
 +
! no. 2 (4 hours)
 +
! no. 1 (6 hours)
 +
! no. 2 (6 hours)
 +
|-
 +
| RBSGFP
 +
| min 0.001
 +
| min 0.001
 +
| 0.021
 +
| 0.004
 +
| 0
 +
| 0
 +
|-
 +
| R0062
 +
| min 0.016
 +
| min 0.011
 +
| 0.001
 +
| 0.020
 +
| 0
 +
| 0
 +
|-
 +
| ptetGFP
 +
| min 0.006
 +
| min 0.004
 +
| 0.011
 +
| min 0.003
 +
| 0
 +
| 0
 +
|-
 +
| F2621GFP
 +
| min 0.014
 +
| min 0.003
 +
| 0.009
 +
| 0.011
 +
| 0
 +
| 0
 +
|-
 +
|}
 +
 +
{|
 +
|  Orange    ||  Apple  ||  more
 +
|-
 +
|  Bread    ||  Pie    ||  more
 +
|-
 +
|  Butter  || Ice cream ||  and more
 +
|}
[https://2011.igem.org/Team:Wageningen_UR/Notebook/Proj1/June Previous Month]
[https://2011.igem.org/Team:Wageningen_UR/Notebook/Proj1/June Previous Month]

Revision as of 13:55, 30 July 2011

Building a Synchronized Oscillatory System

July - Synchronized Oscillatory System

Jul.png

July 13

BioBrick parts were digested using different combinations of restriction enzymes, ligated and run on a gel.


July 18

Gel extracion of the gel run on July 13th was performed. Liquid cultures were made.


July 19

Yesterday's liquid cultures were miniprepped.

July 30

Inoculated 500 mL erlenmeyers with 100 ml LB + 100 ug/ul. E. coli was grown overnight on LB + amp plates and directly used as inoculum. All cultures were done in duplo. E. coli strains used were: ptet-GFP R0062-GFP F2621-GFP RBS-GFP

Cultures were put in a shake incubator at 30°C, 300 rpm. Temperature was set to 30 degrees as GFP is degraded at higher temperatures. The OD600s of every culture were measured 2 hours after inoculation and subsequently each hour, by taking 1 mL samples from the culture. the OD was measured with a Hitachi U1500 spectrophotometer. LB+amp was taken as blank. Fluorescence was measured in a 96-wells plate on a fluorescent plate reader (Molecular Devices, M2 (toxicology). First blank measurements were done with 96 wells plates containing 50 ul LB+amp. The output corresponding with the specific wells was saved and used for further calculations.

OD600 data presented below

{

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