Team:Chiba/Notebook/Calendar/18 September 2009

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([[Team:Chiba/Notebook/Calendar/17_September_2009|17_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/19_September_2009|19_September_2009]])
([[Team:Chiba/Notebook/Calendar/17_September_2009|17_September_2009]] <|>[[Team:Chiba/Notebook/Calendar/19_September_2009|19_September_2009]])
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==LuxR Mutants Sequence==
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 +
Purification of DNA
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*isopropanol precipitation and analysis
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 +
#add isopropanol(75%) 50 uL to PCR solution made yesterday
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#VORTEX and centrifugation(14000 rpm 10 min)
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#put off the supernatant fluid
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#add isopropanol(75%) 50 uL
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#centrifugation(14000 rpm 10 min)
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#put off the supernatant fluid
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#drying 60 degrees C 15min
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#add ddH2O 20 uL and VORTEX
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#sequence analysis
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==Las Check==
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Culture Mix
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#K084007:LasI
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#K084012:LuxI
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#T9002:LuxR-GFP
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#K091134:LasR-GFP
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#I731012:LuxR-RFP
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Sender:Receiver=0.8 mL:0.8 mL
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(1):1 + 4 ---LasI/LasR-GFP
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(2):1 + 3 ---LasI/LuxR-GFP
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(3):1 + 5 ---LasI/LuxR-RFP
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(4):2 + 4 ---LuxI/LasI-GFP
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(5):2 + 3 ---LuxI/LuxR-GFP
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(6):2 + 5 ---LuxI/LuxR-RFP
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(7):4 + flesh LB ---LasR-GFP
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(8):3 + flesh LB ---LuxR-GFP
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(9):5 + flesh LB ---LuxR-RFP
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Result
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:<table width="135" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr align="center">
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:<td width="50">sample</td>
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:<td width="85">fluorescence</td>
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:</tr>
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:<tr>
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:<td align="center">(1)</td>
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:<td align="center">-</td>
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:</tr>
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:<tr>
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:<td align="center">(2)</td>
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:<td align="center">+</td>
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:</tr>
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:<tr>
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:<td align="center">(3)</td>
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:<td align="center">+</td>
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:</tr>
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:<tr>
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:<td align="center">(4)</td>
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:<td align="center">-</td>
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:</tr>
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:<tr>
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:<td align="center">(5)</td>
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:<td align="center">++</td>
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:</tr>
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:<tr>
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:<td align="center">(6)</td>
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:<td align="center">+</td>
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:</tr>
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:<tr>
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:<td align="center">(7)</td>
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:<td align="center">-</td>
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:</tr>
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:<tr>
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:<td align="center">(8)</td>
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:<td align="center">-</td>
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:</tr>
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:<tr>
 +
:<td align="center">(9)</td>
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:<td align="center">+</td>
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:</tr>
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:</table>
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 +
 +
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LasR is Unworking
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== Lab Notebook ==
== Lab Notebook ==
[[Image:Chiba-Labwork-17Sep09.jpg|200px]] [[Image:Chiba-Labwork-18Sep09.jpg|200px]]
[[Image:Chiba-Labwork-17Sep09.jpg|200px]] [[Image:Chiba-Labwork-18Sep09.jpg|200px]]
[[Image:Chiba-Labwork-17Sep09-5.jpg|200px]] [[Image:Chiba-Labwork-17Sep09-2.jpg|200px]]
[[Image:Chiba-Labwork-17Sep09-5.jpg|200px]] [[Image:Chiba-Labwork-17Sep09-2.jpg|200px]]

Latest revision as of 07:25, 23 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

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

Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/18_September_2009"