Team:Chiba/Calendar-Home/3 September 2008

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2 September 2008 <|> 4 September 2008

Contents

Laboratory work

Team:Input

no work

Team:Communication

(2/9)--->Mini prep
  • [http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
  • [http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8)
  • [http://partsregistry.org/Part:BBa_S03154 BBa_S03154]


--->Gel Check
Chiba-0903.JPG
Sample DNA 1
Loading Dye(μL) 1
dH2O(μL) 4
TOTAL(μL) 6
From left;
  • [http://partsregistry.org/Part:BBa_S03154 BBa_S03154] -> OK 33ng/μL
  • [http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7) -> OK
Chiba-0903-2.JPG
From left;
  • [http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8) -> OK
--->Digestion test
  • [http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
  • [http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8)
Digestion SingleDouble
Sample DNA(μL) 13
XbaⅠ(μL) 0.10.1
PstⅠ(μL) 0.10.1
Buffer 2(μL) 0.9-
Buffer 3(μL) --0.8
BSA(μL) 11
dH2O(μL) 75
TOTAL(μL) 1010


--->Gel Check
Chiba-0903-3.JPG
Sample DNA(μL) 10
Loading Dye(μL) 2
TOTAL(μL) 12
From left;
  • Single Digestion : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
  • Single Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK
Chiba-0903-4.JPG
From left;
  • Double Digestion : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
  • Double Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK


(2/9)--->Phenotype-test
  • MIX
  • [http://partsregistry.org/Part:BBa_K084007 BBa_K084007](Plac+RBS+LasI, Competent Cells : JW1908) -> Sample Name : L1~L4
  • [http://partsregistry.org/Part:BBa_K084008 BBa_K084008](Plac+RBS+RhlI, Competent Cells : JW1908) -> Sample Name : R1~R4
  • [http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002](Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)


Sample No. 1234567891011
L1 2mL----------
L2-2mL---------
L3--2mL--------
L4---2mL-------
R1----1mL------
R2-----1mL-----
R3------1mL----
R4-------1mL---
[http://partsregistry.org/Part:BBa_S03154 BBa_S03154]--------2mL--
AHL(100μM)----------1μL
[http://partsregistry.org/Part:BBa_T9002　 BBa_T9002]2mL2mL2mL2mL1mL1mL1mL1mL2mL1mL1mL
IPTG(100μM)1μL1μL1μL1μL1μL1μL1μL1μL---
  • Incubated for 8hours at 37 degrees
  • Spindown (max rpm, 3 min)
  • The measurement of the intensity GFP(visual judgment)
Sample No. 1234567891011
the intensity GFP +++++++++++++-


  • Left for 12 at room temperture.
  • Resuspensioned
  • measured the intensity of GFP by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6)
Sample No. 1234567891011
the intensity GFP 25.6518.9120.3921.8526.6226.5830.0733.6027.4110.7255.54

Team:Output

Digestion test

  • [http://partsregistry.org/Part:BBa_R0079 BBa_R0079]①
  • [http://partsregistry.org/Part:BBa_R0071 BBa_R0071]②
  • [http://partsregistry.org/Part:BBa_R0077 BBa_R0077]③
  • [http://partsregistry.org/Part:BBa_R0078 BBa_R0078]④
  • [http://partsregistry.org/Part:BBa_R0062 BBa_R0062]⑤
Sample No. ①~⑤
DNA tamplate(μL) 5
Buffer3(μL) 1
dH2O(μL) 4
EcoRⅠ(μL) 0.1
TOTAL(μL) 10

-->37℃30min

-->Gel check

Sample No. ①~⑤
DNA(digested sample(μL)) 10
loading Dye(μL) 2
TOTAL(μL) 12
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