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Team:Chiba/jk/γ/week 5
From 2008.igem.org
Contents |
Week 5
14,September,2008
Digestion test
BBa_Q04510
BBa_I716463
BBa_C0051
BBa_C0070
BBa_J06620
BBa_J22141
BBa_C0076
BBa_I716542
BBa_I716462
BBa_C0078
BBa_J01101
BBa_C0071
BBa_I712019
BBa_S01416
BBa_C0079
BBa_C0077
BBa_J23007
BBa_I7100(x2)
BBa_S01416
| Sample No | each |
| DNA(digestion sample) | 3 |
| Buffer2(x10) | 1 |
| dH2O | 6 |
| EcoRⅠ | 0.075 |
| TOTAL | 10 |
->37℃ 1hour
Gel cheack
| Sample No | each |
| DNA(digestion sample) | 10 |
| Loading Dye | 2 |
| TOTAL | 12 |
16,September,2008
Digestion test
BBa_R0040(ptet)+BBa_S03158(RBS+RhlR)(x2)
BBa_R0040(ptet)+BBa_S03156(RBS+lasR)(x5)
BBa_R0040(ptet)+BBa_S03160(RBS+cinR)(x2)
BBa_S03158(RBS+RhlR)
BBa_S03156(RBS+lasR)
BBa_S03160(RBS+cinR)
| Sample No | each |
| DNA | 3 |
| EcoRⅠ | 0.083 |
| PstⅠ | 0.083 | BSA(x10) | 1 |
| Buffer3(x10) | 1 |
| dH2O | 5 |
| TOTAL | 10 |
->37℃ 1hour
Gel cheack
| Sample No | each |
| DNA | 10 |
| Loading Dye | 2 |
| TOTAL | 12 |
TRANSFORATION
BBa_J37019(LuxI+RBS+LuxR)->no colony
BBa_J37003(RBS+LuxR)
BBa_J37020(LuxI+LuxR+pLux+GFP)
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&insert | Vector |
| DNA(vector) | 3.5 | 3.5 |
| DNA(insert) | 7.5 | - |
| Ligase | 1 | 1 |
| Ligase Buffer | 4 | 4 |
| dH2O | 4 | 4 |
| TOTAL | 20 | 20 |
TRANSFORMATION
BBa_J54103(RBS+GFP)
BBa_P2006
Gel Cheack
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)
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