Template:Kyoto/Notebook/Sep 6

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Contents

Sep 6

Liquid Culture

No name

Samplemedium
9/4 RBS-Lysis3+DT3Plusgrow medium(+Cp)
9/4 RBS-Lysis1+DT3Plusgrow medium(+Cp)
9/4 PSB4K5Plusgrow medium(+K) 

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

Miniprep

No Name

DNAconcentration [µg/mL]260/280260/230
Plac153.72.032.10
Pλ-luxⅠ-(1)76.52.072.06
Pλ-luxⅠ-(2)86.42.082.10
DT166.21.652.12
J23100190.11.682.34

Restriction Enzyme Digestion

Tatsui

9/6 RBS-lysis2-DT (78.3 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts22µL0µL0µL1µL1µL3µL3µL0µL30µL
NC1.3µL0µL0µL0µL0µL1µL1µL6.7µL10µL
  • at 37°C, for 1h
aptamer-DT (251.7 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts0.4µL0µL0µL1µL1µL3µL3µL14.1µL30µL
NC0.4µL0µL0µL0µL0µL1µL1µL7.6µL10µL
  • at 37°C, for 1h
Pλ-luxI (86.4 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts23.1µL1µL1µL0µL0µL3µL3µL1.9µL30µL
NC1.2µL0µL0µL0µL0µL1µL1µL6.8µL10µL
  • at 37°C, for 1h

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder--
28/20 RBS-lysis1-(1)EcoRI & SpeI
3
4----
58/28 RBS-GFP-DT-(2)XbaI & PstI
6

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

LaneDNAEnzyme
1100bp ladder--
28/28 PluxSpeI & PstI
3
4----
58/20 RBS-tetR-DT-(1)XbaI & PstI
6

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

Nameconcentration[µg/mL]260/280260/230
Pcon-RBS-GFP-DT (EcoRI&SpeI)7.11.880.07
Pcon-RBS-luxR-DT (EcoRI&XbaI)15.31.930.70
Plux (SpeI&PstI)4.51.790.31
RBS-GFP-DT (XbaI&PstI)6.02.160.03
RBS-tetR-DT (XbaI&PstI)7.61.910.38


Liquid Culture

No name

Samplemedium
8/21 RBS-Lysis2-9Plusgrow medium(+Amp)
8/21 RBS-Lysis2-12Plusgrow medium(+Amp)
8/21 RBS-Lysis2-14Plusgrow medium(+Amp) 

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png