Team:Chiba/Calendar-Home/22 August 2008

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===Team:Input===
===Team:Input===
'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]''':
'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]''':
-
Minipreped the cultures(->21/8).
+
Minipreped the cultures(<--21/8).
*[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
*[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
Line 19: Line 19:
insert check:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
insert check:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
-
*[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
+
#[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
-
*[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
+
#[http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
-
*[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
+
#[http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
-
*[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)
+
#[http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)
-
*[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
+
#[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
-
*[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
+
#[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
 +
 
 +
 
-
[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)と[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)はsingleのみ、ほかはsingleとdoubleの両方。
 
-
<BR>
 
-
<BR>混ぜ表
 
<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
-
<td width="257"></td>
+
<td width="257" >Sample No.</td>
-
<td>double</td><td>simgle</td>
+
<td>1-4</td><td>1-6</td>
<tr>
<tr>
-
<td>dH<sub>2</sub>O</td>
+
<td>dH<sub>2</sub>O(&mu;l)</td><td>14</td><td>25</td>
-
<td>14</td><td>25</td>
+
</tr>
</tr>
<tr>
<tr>
-
<td>10×BSA</td>
+
<td>10×BSA(&mu;l)</td>
<td>7</td><td>10</td>
<td>7</td><td>10</td>
</tr>
</tr>
<tr>
<tr>
-
<td>10×NE</td>
+
<td>10×NE(&mu;l)</td>
<td>7</td><td>10</td>
<td>7</td><td>10</td>
</tr>
</tr>
<tr>
<tr>
-
<td>EcoRI</td>
+
<td>EcoRI(&mu;l)</td>
<td>3.5</td><td>5</td>
<td>3.5</td><td>5</td>
</tr>
</tr>
<tr>
<tr>
-
<td>PstI</td>
+
<td>PstI(&mu;l)</td>
<td>3.5</td><td>-</td>
<td>3.5</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>Total(&mu;l)</td>
 +
<td>35</td><td>50</td>
</tr>
</tr>
</table>
</table>
-
<BR>UV照射実験(液体編)(~24)
 
-
<BR>-Ptrc-LuxR-Plux-cI-colE1-Amp-,-PcI-GFP-p15a-Cm-(BW)の2plasmidを使用。
 
-
<BR>グリストをつついて2ml培養×2(LB-Amp,Cm)(LB-Amp,cm,AHL100nM)
 
-
<BR>37℃で、12h培養後、濁ったらそのまま小さなプレートにLB培地ごと移してUVを照射する。UVの波長は254nm,UVランプまでの距離は8cm。
 
-
<BR>UVを照射して、15min,4h,8h,12h,たったら、両方のプレートからLB培地ごと20μlずつ採取しそれぞれ10<sup>5</sup>希釈し、そこから20μlをそれぞれ(LB-Amp,Cm)(LB-Amp,cm,AHL100nM)のプレートに撒いた。
 
-
<BR>37℃で12h培養した後のコロニー数
+
UV irradiation test
 +
 
 +
* two plasmids from
 +
          o -Ptrc-LuxR-Plux-cI-colE1-Amp-
 +
          o -PcI-GFP-p15a-Cm-(JW1908)
 +
#Incubated cultures(from Glycerol Stocks) with 2mL of LB-Ampicillin, Chloramphenicol Medium and LB-Ampicilline, Chloramphenicol and AHL100nm Medium for 12 hours at 37 degree.
 +
#moved the cultures to small plates and started UV-irradiation. (wavelength:254nm,distance from the UV lamp to the cultures were 8cm.)
 +
#After exposing both plates to UV light for 15min, 4h, 8h, or 12h, we took 20ul from each plate, diluted 10<sup>5</sup>-fold, and plated 20ul of the resulting solution t (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates.
 +
 
 +
 
 +
<BR>The number of colonies aftetr culturing at 37&deg;C for 12h.
<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<table width="200" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
<td width="257"></td>
<td width="257"></td>
-
<td>AHLなし</td><td>AHL100nM</td>
+
<td>no AHL</td><td>AHL100nM</td>
<tr>
<tr>
<td>15min</td>
<td>15min</td>
-
<td>136</td><td>0?</td>
+
<td>136</td><td>0</td>
</tr>
</tr>
<tr>
<tr>
Line 81: Line 88:
</tr>
</tr>
</table>
</table>
 +
<BR>   
 +
result
<BR>
<BR>
-
いづれのプレートもAHLが入っていないものはGFPが確認できたが、AHL100nMのものは確認できなかった。
+
GFP fluorescence was observed from plates without AHL but not from
 +
plates containing 100nM AHL.
===Team:Communication===
===Team:Communication===
:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
:competent cells : XL10G
:competent cells : XL10G
-
::*BBa_J13002(2007) [http://partsregistry.org/Part:BBa_J13002]
+
::*[http://partsregistry.org/Part:BBa_J13002　 BBa_J13002](2007)
-
::*BBa_J13002(2006) [http://partsregistry.org/Part:BBa_J13002]
+
::*[http://partsregistry.org/Part:BBa_J13002　 BBa_J13002](2006)
-
::*BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
+
::*[http://partsregistry.org/Part:BBa_J04500　 BBa_J04500](2007)
-
::*BBa_J04500(2006) [http://partsregistry.org/Part:BBa_J04500]
+
::*[http://partsregistry.org/Part:BBa_J04500　 BBa_J04500](2006)
:(Locate & 2 Punch ~iyama ver.~)
:(Locate & 2 Punch ~iyama ver.~)
-
::*BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates  
+
::*[http://partsregistry.org/Part:BBa_C0076　 BBa_C0076](2008)  -> No colonies on plates  
-
::*BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates  
+
::*[http://partsregistry.org/Part:BBa_C0077 BBa_C0077](2008)　-> No colonies on plates  
-
::*BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
+
::*[http://partsregistry.org/Part:BBa_C0078　 BBa_C0078](2008)  -> No colonies on plates
-
::*BBa_J04500(2008) [http://partsregistry.org/Part:BBa_J04500] -> No colonies on plates
+
::*[http://partsregistry.org/Part:BBa_J04500  BBa_J04500](2008) -> No colonies on plates
--->(24/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
--->(24/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
-
 
===Team:Output===
===Team:Output===
[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
-
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](Luciferase)
+
*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]
*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]
-
*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202](LacZ)
+
*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]
*PUC19
*PUC19
*PGEX VENUS YFP(GST fusion)
*PGEX VENUS YFP(GST fusion)

Latest revision as of 03:02, 30 October 2008

>Home | Notebook

21 August 2008 <|> 23 August 2008

Contents

Laboratory work

Team:Input

Mini prep: Minipreped the cultures(<--21/8).

  • [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
  • [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
  • [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
  • [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
  • [http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
  • [http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)


insert check:Digestion

  1. [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
  2. [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
  3. [http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
  4. [http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)
  5. [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
  6. [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)


Sample No. 1-41-6
dH2O(μl)1425
10×BSA(μl) 710
10×NE(μl) 710
EcoRI(μl) 3.55
PstI(μl) 3.5-
Total(μl) 3550


UV irradiation test

  • two plasmids from 
         o -Ptrc-LuxR-Plux-cI-colE1-Amp-
         o -PcI-GFP-p15a-Cm-(JW1908)
  1. Incubated cultures(from Glycerol Stocks) with 2mL of LB-Ampicillin, Chloramphenicol Medium and LB-Ampicilline, Chloramphenicol and AHL100nm Medium for 12 hours at 37 degree.
  2. moved the cultures to small plates and started UV-irradiation. (wavelength:254nm,distance from the UV lamp to the cultures were 8cm.)
  3. After exposing both plates to UV light for 15min, 4h, 8h, or 12h, we took 20ul from each plate, diluted 105-fold, and plated 20ul of the resulting solution t (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates.



The number of colonies aftetr culturing at 37°C for 12h.

no AHLAHL100nM
15min 1360
4h 4031
8h 100
12h 00


result
GFP fluorescence was observed from plates without AHL but not from plates containing 100nM AHL.

Team:Communication

Transformation
competent cells : XL10G
  • [http://partsregistry.org/Part:BBa_J13002　 BBa_J13002](2007)
  • [http://partsregistry.org/Part:BBa_J13002　 BBa_J13002](2006)
  • [http://partsregistry.org/Part:BBa_J04500　 BBa_J04500](2007)
  • [http://partsregistry.org/Part:BBa_J04500　 BBa_J04500](2006)
(Locate & 2 Punch ~iyama ver.~)
  • [http://partsregistry.org/Part:BBa_C0076　 BBa_C0076](2008) -> No colonies on plates
  • [http://partsregistry.org/Part:BBa_C0077 BBa_C0077](2008)　-> No colonies on plates
  • [http://partsregistry.org/Part:BBa_C0078　 BBa_C0078](2008) -> No colonies on plates
  • [http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2008) -> No colonies on plates


--->(24/8)Mini prep

Team:Output

Mini prep

  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]
  • [http://partsregistry.org/Part:BBa_J33202 BBa_J33202]
  • PUC19
  • PGEX VENUS YFP(GST fusion)


Transformation

  • PGEX VENUS YFP(GST fusion)
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