Team:Chiba/Calendar-Home/21 August 2008

From 2008.igem.org

(Difference between revisions)
(Team:Input)
(Team:Input)
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*UV irradiation test  (plate version)
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*UV irradiation test  (plate phase)
*two plasmids from ---
*two plasmids from ---
**(Repressor)-Ptrc-LuxR-Plux-cI-colE1-Amp-
**(Repressor)-Ptrc-LuxR-Plux-cI-colE1-Amp-

Revision as of 01:08, 30 October 2008

>Home | Notebook

20 August 2008 <|> 22 August 2008

Contents

Laboratory work

Team:Input

(-->20/8)

Inoculated transformants for 12 hours in LB 2mL containing Ampicillin.

  • [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)

BBa_J22141:After inoculated for 11 hours, added 20μl IPTG and cultured one hour.


  • UV irradiation test (plate phase)
  • two plasmids from ---
    • (Repressor)-Ptrc-LuxR-Plux-cI-colE1-Amp-
    • (Reporter)-PcI-GFP-p15a-Cm-(BW)
  1. Inoculated (Reporter) culture from glycerol stock(-80°C freezer) in 2mL LB medium.(LB-Amp,Cm)(LB-Amp,Cm,AHL100nM)


Inoculated transformants for 12 hours in 2ml LB-Ampicillin, Chloramphenicol medium and LB-Ampicillin, Chloramphenicol, 100nmAHL medium .(37&ged;C, 12h). それぞれを105希釈してAHLが入っていないものはLB-Amp,Cm,AHLが入っているものはAHL-Amp,Cm,AHL100nMのプレートにそれぞれ20μl撒く。
37°Cで12h培養後、きちんとコロニーができていることを確認し、両方のプレートにUVを照射する。UVの波長は254nm,UVランプとプレートの距離は14cm。
UVを照射して9h後、21h後にそれぞれ両方のプレートに次の同じ操作をする。
両方のプレートのコロニーをつついてそれぞれ(LB-Amp,Cm),(LB-Amp,Cm,AHL100nM)2ml培養する。
37°C,で12h培養後、濁っていたので、それぞれを(LB-Amp,Cm),(LB-Amp,Cm,AHL100nM)のプレートに105希釈して20μl撒いた。


  • Colony count
no AHLAHL100nM
9h 606453
12h 146151

Colonies of no AHL plate were AHLが入っていないほうのコロニーはいづれも光っていた。→機能チェックはOK

Team:Communication

(20/8)--->PCR
  • [http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2007)
  • [http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2006)
  • [http://partsregistry.org/Part:BBa_C0076 BBa_C0076](2007)
  • [http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2007)
  • [http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2006)


DNA Template 1
dNTP mix 10
Foward Primer 5
Reverse Primer 5
DNA polymerase TAQ 1
Thermopol Buffer 5
dH2O 28
TOTAL 50μL


95°C,5min -> ( 95°C,1min -> 52°C,1min -> 72°C,1min )・・・25cycles -> 72°C,10min -> 6°C


--->Gel Check
Chiba-0821.JPG
Sample DNA 3
Loading Dye 2
dH2O 7
TOTAL 12μL
From left;
  • [http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2007) -> None
  • [http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2006) -> None
  • [http://partsregistry.org/Part:BBa_C0076 BBa_C0076](2007) -> None
  • [http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2007) -> OK
  • [http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2006) -> OK


Transformation
competent cells : XL10G
  • [http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2007)
  • [http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2006)
  • [http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2007)
  • [http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2006)
  • [http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2007)
  • [http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2006)



--->(23/8)Digestion

Team:Output

Transformation

  • [http://partsregistry.org/Part:BBa_J32007 BBa_J32007](2007)-->no colony-->no colony(2 times)-->colony(3times)
  • [http://partsregistry.org/Part:BBa_B0034 BBa_B0034](2007)-->colony