Template:Kyoto/Notebook/Sep 15

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Contents

Sep 15

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
Pλ-luxI225 1.59 0.83

Liquid Culture

No name

Samplemedium
9/13 Pλ-luxI-1
9/7 pSB4K5-1
9/11 Plac-3

Ligation

じっけんしゃ

stateVectorInserterLigation High ver.2
experiment9/13 Pcon(SpeI & PstI)3.6 µL9/12 RBS-lacZα-DT(XbaI & PstI)4.7 µL8 µL

Restriction Enzyme Digestion

No Name

DNAEcorISpeIBufferBSAMilliQtotal
9/7Pcon-RBS-tetR-DT (EcoRI & SpeI)15 µL1 µL1 µL3 µL3 µL6 µL30 µL
NC0.7 µL0 µL0 µL1 µL1 µL7.3 µL10 µL


DNAEcoRISpeIBufferBSAMilliQtotal
9/12 Pcon-pT181 attenuator-DT (EcoRI & SpeI)6.8 µL1 µL1 µL3 µL3 µL15.2 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAEcoRISpeIBufferBSAMilliQtotal
9/12 Pcon-apt12_1R-DT(EcoRI & SpeI)6.6 µL1 µL1 µL3 µL3 µL15.4 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAEcoRISpeIBufferBSAMilliQtotal
9/8 Pcon-spin-DT (EcoRI & SpeI)7.6 µL1 µL1 µL3 µL3 µL15.4 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
9/11 Pcon-pT181 anticodone-spin-DT(XbaI & PstI)7.2 µL1 µL1 µL3 µL3 µL14.8 µL30 µL
NC0.4 µL0 µL0 µL1 µL1 µL7.6 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
8/17 RBS-lacZα-DT (XbaI & PstI)5.3 µL1 µL1 µL3 µL3 µL16.7 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL


DNAEcoRIXbaIBufferBSAMilliQtotal
9/7 Ptet-RBS-LacZα-DT (EcoRI & XbaI)2.9 µL1 µL1 µL3 µL3 µL18.5 µL30 µL
NC1.5 µL0 µL0 µL1 µL1 µL7 µL10 µL


Electrophoresis

No Name

LaneSampleEnzyme1Enzyme2
1,14
2 Pcon-RBS-tetR-DT EcoRI SpeI
3 Pcon-RBS-tetR-DT
4 Pcon-pT181 attenuator-DT EcoRI SpeI
5 Pcon-pT181 attenuator-DT
6 Pcon-apt12-1R-DT EcoRI SpeI
7 Pcon-apt12-1R-DT
8 Pcon-spin-DT EcoRI SpeI
9 Pcon-spin-DT
10 Pcon-pT181 antisense-spin-DT XbaI PstI
11 Pcon-pT181 antisense-spin-DT
12 RBS-lacZα-DT XbaI PstI
13 RBS-lacZα-DT
15 Ptet-RBS-lacZα-DT EcoRI XbaI
16 Ptet-RBS-lacZα-DT
17 1kbp ladder

File:Igku xxxxxx.xxx

Gel Extraction

No Name 1

LaneDNAEnzyme
1 100bp ladder
2~4 Pcon-pT181 attenuator-DT EcoRI & SpeI
6~8 Pcon-apt12_1R-DT
10~12 Pcon-spin-DT

2

LaneDNAEnzyme
1 100bp ladder
2~4 Pcon-pT181 anticodone-spin-DT XbaI & PstI
6~8 RBS-lacZα-DT XbaI & PstI
10~12 Pcon-RBS-tetR-DT EcoRI & SpeI

File:Igku xxbeforexx.xxx
File:Igku xxafterxx.xxx

Nameconcentration[µg/mL]260/280260/230
9/15 Ptet-RBS-lacZα-DT (EcoRI & SpeI) 1.9 1.08 0.77
9/15 Pcon-RBS-tetR-DT (EcoRI & SpeI) 5.9 1.30 0.17
9/15 RBS-lacZα-DT (XbaI & PstI) 1.0 0.85 0.44
Pcon-pT181 anticodone-spin-DT (XbaI & PstI) 1.3 0.95 0.28
Pcon-spin-DT 0.9 1.37 0.05
Pcon-apt12_1R-DT 1.7 1.42 0.29
Pcon-pT181 attenuator-DT (EcoRI & SpeI) 2.1 1.53 0.46

Ligation

No Name

stateVectorInserterLigation High ver.2
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-RBS-tetR-DT EcoRI & SpeI1.9 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-spin-DT EcoRI & SpeI4.7 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-apt12_1R-DT EcoRI & SpeI2.0 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-pT181 attenuator-DT EcoRI & SpeI3.0 µLxx µL
experiment9/13 pSB1C3 XbaI & PstI1.9 µL9/15 Pcon-pT181 anticodone-spin-DT XbaI & PstI4.4 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/3 pT181 attenuator EcoRI & SpeIxx µL3.8 µL
experiment9/6 Pcon SpeI & PstI1.4 µL9/15 RBS-lacZα-DT XbaI & PstIxx µL2.0 µL
experiment9/14 pSB 4k5 EcoRI & SpeI µL9/15 Pcon-spin-DT EcoRI & SpeIxx µL9.4 µL
experiment9/15 Peet-RBS-lacZα-DT EcoRI & XbaI2.6 µLPcon-RBS-tetR-DT EcoRI & SpeI1.5 µLxx µL