Team:Chiba/jk/γ/week 5

From 2008.igem.org

実験ログ

Contents

Week 5

14,September,2008


Time Responce Liquid
Digestion test

  • [http://partsregistry.org/Part:BBa_Q04510 BBa_Q04510]
  • [http://partsregistry.org/Part:BBa_I716463 BBa_I716463]
  • [http://partsregistry.org/Part:BBa_C0051 BBa_C0051]
  • [http://partsregistry.org/Part:BBa_C0070 BBa_C0070]
  • [http://partsregistry.org/Part:BBa_J06620 BBa_J06620]
  • [http://partsregistry.org/Part:BBa_J22141 BBa_J22141]
  • [http://partsregistry.org/Part:BBa_C0076 BBa_C0076]
  • BBa_I716542
  • [http://partsregistry.org/Part:BBa_I716462 BBa_I716462]
  • [http://partsregistry.org/Part:BBa_C0078 BBa_C0078]
  • [http://partsregistry.org/Part:BBa_J01101 BBa_J01101]
  • [http://partsregistry.org/Part:BBa_C0071 BBa_C0071]
  • [http://partsregistry.org/Part:BBa_I712019 BBa_I712019]
  • [http://partsregistry.org/Part:BBa_S01416 BBa_S01416]
  • [http://partsregistry.org/Part:BBa_C0079 BBa_C0079]
  • [http://partsregistry.org/Part:BBa_C0077 BBa_C0077]
  • [http://partsregistry.org/Part:BBa_J23007 BBa_J23007]
  • [http://partsregistry.org/Part:BBa_I7100 BBa_I7100](x2)
  • [http://partsregistry.org/Part:BBa_S01416 BBa_S01416]
Sample No each
DNA(digested sample) 3
Buffer2(x10) 1
dH2O 6
EcoRⅠ 0.075
TOTAL 10

-->37℃ 1hour

Gel check

Sample No each
DNA(digestion sample) 10
Loading Dye 2
TOTAL 12

16,September,2008


Digestion test

  • [http://partsregistry.org/Part:BBa_R0040 BBa_R0040](ptet)+[http://partsregistry.org/Part:BBa_S03158 BBa_S03158](RBS+RhlR)(x2)
  • [http://partsregistry.org/Part:BBa_R0040 BBa_R0040](ptet)+[http://partsregistry.org/Part:BBa_S03156 BBa_S03156](RBS+lasR)(x5)
  • [http://partsregistry.org/Part:BBa_R0040 BBa_R0040](ptet)+[http://partsregistry.org/Part:BBa_S03160 Ba_S03160](RBS+cinR)(x2)
  • [http://partsregistry.org/Part:BBa_S03158 BBa_S03158](RBS+RhlR)
  • [http://partsregistry.org/Part:BBa_S03156 BBa_S03156](RBS+lasR)
  • [http://partsregistry.org/Part:BBa_S03160 Ba_S03160](RBS+cinR)

</tr>

Sample No. each
DNA 3
EcoRⅠ 0.083
PstⅠ 0.083
BSA(x10) 1
Buffer3(x10) 1
dH2O 5
TOTAL 10

-->37℃ 1hour

Gel check

Sample No. each
DNA 10
Loading Dye 2
TOTAL 12

Transformation

  • [http://partsregistry.org/Part:BBa_J37019 BBa_J37019](LuxI+RBS+LuxR)->no colony
  • [http://partsregistry.org/Part:BBa_J37033 BBa_J37033](RBS+LuxR)
  • [http://partsregistry.org/Part:BBa_J37020 BBa_J37020](LuxI+LuxR+pLux+GFP)
  • [http://partsregistry.org/Part:BBa_J37032 BBa_J37032](pLux+GFP)

PCR

  • Venus YFP(ptet+LuxR+YFP)
Sample No. YFP
DNA 1
Venus Fwd 5
Rev (VR) 5
Thermo pol Buffer 10
dNTP 10
Vent 1
dH2O 68
TOTAL 100

17,September,2008

Transformation

  • BBa_I13500(RBS+GFP)
  • BBa_J37019(Luxpr+LuxR)-->no colony


MINI Prep

  • BBa_J37033(RBS+LuxR)
  • BBa_J37020(Luxpr+LuxR+LuxR+GFP)
  • BBa_J37032(pLux+GFP)


Digestion

  • BBa_I7100(ptetR+RBS)
  • BBa_S03119(ptet+LuxR)
  • Ligation product(ptet+LasR)
  • Ligation product(ptet+RhlR)
  • Ligation product(ptet+cinR)
Sample No each
DNA template 25
EcoRⅠ 1
PstⅠ 1
BSA(x10) 4
EcoRⅠ Buffer 4
dH2O 5
TOTAL 40


-->37℃ 1hour
->Next day


18,September,2008


SAP 1μl
->37℃ 1hour
->30min 65℃
Zymo ->25μl
Ligation
vector:BBa_I7100
insert:BBa_S03119(ptet+LuxR),Ligation product(ptet+LasR,ptet+RhlR,ptet+cinR)

Sample No Vector&insertVector
DNA(vector) 3.53.5
DNA(insert) 7.5-
Ligase 11
Ligase Buffer 44
dH2O 44
TOTAL 2020


TRANSFORMATION
BBa_J54103(RBS+GFP)
BBa_P2006

Gel check
Ligation product(Venus YFP)
BBa_J37033
BBa_J37020
BBa_J37032

Sample No each
DNA each
Loading Dye 1
dH2O 4
TOTAL 6


19,September,2008


colony PCR
ptet+LuxR(x16)
ptet+LasR(x7)
ptet+RhlR(x5)
ptet+cinR(x5)


20,September,2008


>Last Week
>Next Week


ホーム メンバー紹介 プロジェクト紹介 Parts Submitted to the Registry モデリング ノート