Team:Chiba/Notebook/Calendar/24 September 2009

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Revision as of 04:46, 26 September 2009

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(23_September_2009 <|>25_September_2009)

Examine limit of AHL generation(1)-3

Yesterday's operation is here.

Today's operation

14:00

We made plates from each of mixture and 10 mL of LB-Amp, Cm solution medium.

Element of mixtures

Sample Namber	1	2	3	4	5	6	7	8
supernatant solution	5 mL	---	---	---	---	---	---	20 μL
1	---	5 mL	---	---	---	---	---	---
2	---	---	5 mL	---	---	---	---	---
3	---	---	---	5 mL	---	---	---	---
4	---	---	---	---	5 mL	---	---	---
5	---	---	---	---	---	5 mL	---	---
6	---	---	---	---	---	---	5 mL	---
LB-Amp,Cm	10 mL	10 mL	10 mL	10 mL	10 mL	10 mL	10 mL	9.98 mL
Total	10 mL	10 mL	10 mL	10 mL	10 mL	10 mL	10 mL	10 mL

Transformation(2)-2

Yesterday's operation is here.

Today's operation

11:15-

We picked colony(plux-GFP only) and cultured it.

15:50-

Main culture

20:30-

Mini prep.

And resultant DNA was saved in freezer.

Transformation(3)-1

Plasmids

ptet-GFP pSB1A2 [http://partsregistry.org/Part:BBa_I13522 BBa_I13522](Amp)

ptet-RFP pSB1A3 [http://partsregistry.org/Part:BBa_I13521 BBa_I13521](Amp)

ptet-CFP pSB1A2 [http://partsregistry.org/Part:BBa_I13600 BBa_I13600](Amp)

mRFP without Nco1 site(Cm)

Competent Cells

JW1262

21:50-

We cultured each plates.

Digestion Test

Samples

1 : ECFP([http://partsregistry.org/Part:BBa_I6057 BBa_I6057])

2 : mCherry([http://partsregistry.org/Part:BBa_E2060 BBa_E2060])

3 : V-YFP([http://partsregistry.org/Part:BBa_K084003 BBa_K084003])

4 : mOrange([http://partsregistry.org/Part:BBa_E2050 BBa_E2050])

5 : LacZ α([http://partsregistry.org/Part:BBa_T9003 BBa_T9003])

6 : mRFP without Nco1 site

Doble Digestion

Sample	3 μL
EcoR1	0.20 μL
Pst1	0.20 μL
Buffer	1 μL
BSA	1 μL
dw	4.6 μL
Total	10 μL

Doble Digestion's Master Mix

Sample	3 μL
EcoR1	1.4 μL
Pst1	1.4 μL
Buffer	7 μL
BSA	7 μL
dw	32.2 μL
Total	49 μL

Single Digestion's Master Mix

EcoR1	1.4 μL
Buffer	7 μL
dw	40.6 μL
Total	49 μL

Doble Digestion

Sample	3 μL
EcoR1	0.20 μL
Pst1	0.20 μL
Buffer	1 μL
BSA	1 μL
dw	4.6 μL
Total	10 μL

12:50

Warm up at 37 degrees Celsius

13:30

Gel electrophoretic analysis

To judge character of LuxR mutants(3)-1

We poured 1 mL of LB-Amp, Cm liquid medium in 96 deep well and added glycerol stocks.

22:10-

We cultured and shook it at 37 degrees Celsius.

@@ Line 115: / Line 115: @@
 <td>10 mL</td>
 <td>10 mL</td>
-<td>9.8 mL</td>
+<td>9.98 mL</td>
 </tr>
 <tr>