Template:Kyoto/Notebook/Sep 17

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===PCR===
===PCR===
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<div class="experiment">
<div class="experiment">
<span class="author">Hirano</span>
<span class="author">Hirano</span>
{| class="wikitable"
{| class="wikitable"
-
!Pcon-tet ap-DT(VF)||BigDye||5x buffer||primer(VF)||MilliQ||total
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!Pcon-tetR aptamer-DT(VF)||BigDye||5x buffer||primer(VF)||MilliQ||total
|-
|-
|1.3||0.5||1.75||0.5||0.45||10.5
|1.3||0.5||1.75||0.5||0.45||10.5
|}
|}
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{| class="wikitable"
 
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!PreDenature||Denature||Annealing||Extension||cycle
 
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|-
 
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|96&deg;C||96&deg;C||50&deg;C||60&deg;C||--
 
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|-
 
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|2m||10s||5s||4m||30cycles
 
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|}
 
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</div>
 
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<div class="experiment">
 
{| class="wikitable"
{| class="wikitable"
!Pcon-tet ap-DT(VR)||BigDye||5x buffer||primer(VR)||MilliQ||total
!Pcon-tet ap-DT(VR)||BigDye||5x buffer||primer(VR)||MilliQ||total
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!PreDenature||Denature||Annealing||Extension||cycle
!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
-
|96&deg;C||96&deg;C||50&deg;C||60&deg;C||--
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|96 &deg;C||96 &deg;C||50 &deg;C||60 &deg;C||--
|-
|-
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|2m||10s||5s||4m||30cycles
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|2 min||10 sec||5 sec||4 min||30 cycles
|}
|}
</div>
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===Electrophoresis===
===Electrophoresis===
<div class="experiment">
<div class="experiment">

Revision as of 11:54, 24 September 2013

Contents

Sep 17

Liquid Culture

Tatsui and Hirano

Samplemedium
Spinach-DTLB medium(+CP)
Pcon-Spinach-DTLB medium(+Amp)　
Pcon-pT181 antisense-Spinach-DTLB medium(+Amp)　

PCR

Hirano

Pcon-tetR aptamer-DT(VF)BigDye5x bufferprimer(VF)MilliQtotal
1.30.51.750.50.4510.5
Pcon-tet ap-DT(VR)BigDye5x bufferprimer(VR)MilliQtotal
1.30.51.750.50.4510.5
PreDenatureDenatureAnnealingExtensioncycle
96 °C96 °C50 °C60 °C--
2 min10 sec5 sec4 min30 cycles

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
11kbpladder----
2Pcon-RBS-tetR-DTXbaIPstI
3Pcon-RBS-tetR-DT----
4pSB4K5X7EcoRIXbaI
5pSB4K5X7----
6PSB1C3EcoRISpeI
7PSB1C3EcoRIPstI
8PSB1C3----
9pT181attenuator----
10pT181attenuator----
111kbpladder----

File:Igku xxxxxx.xxx

Ligation

No Name

stateVectorInserterLigation High ver.2
experiment8/28 Plux-(SpeI&PstI)5.0µg9/13 RBS-lysis1-DT-(XbaI&PstI)3.5µg3.5µg
experiment8/28 Plux-(SpeI&PstI)5.0micro;g9/13 RBS-lysis2-DT-(XbaI&PstI)3.6µg3.5µg
experiment8/28 Plux-(SpeI&PstI)11.0µg9/13 RBS-lysis3-DT-(XbaI&PstI)9.5µg3.5µg
experiment9/14 Plac-(SpeI&PstI)2.4µg9/16 PT181-attenuator-DT-(XbaI&PstI)4.8µg3.5µg
experiment9/14 Plac-(SpeI&PstI)2.4µg9/3 PT181-antisense-(XbaI&PstI)2.0µg2.2µg
experiment9/14 Plac-(SpeI&PstI)2.4µg9/10 aptamer12-1R-DT-(XbaI&PstI)1.8µg2.1µg
experiment9/13 pSB1C3-(XbaI&PstI)2.0µg9/15 Pcon-PT181 antisense-spinach-DT-(XbaI&PstI)12.0µg3.5µg
experiment9/16 pSB1C3-(EcoRI&SpeI)1.0µg9/15 Pcon-spinach-DT-(EcoRI&SpeI)4.1µg2.0µg
experiment9/16 pSB1C3-(EcoRI&SpeI)1.0µg9/15 Pcon-RBS-tetR-DT-(EcoRI&SpeI)2.0µg1.5µg
experiment9/16 pSB1C3-(EcoRI&SpeI)1.0µg9/15 Pcon-PT181-attenuator-DT-(EcoRI&SpeI)2.4µg1.7µg
experiment9/16 pSB1C3-(EcoRI&SpeI)1.0µg9/15 Pcon-aptamer12-112-DT-(EcoRI&SpeI)17.0µg1.5µg
experiment9/16 pSB1C3-(XbaI&PstI)1.0µg9/15 Pcon-aptamer12-112-DT-(EcoRI&SpeI)4.4µg3.5µg

Gel Extraction

No Name

LaneDNAEnzyme
11kbpladder--
2Pt181 attenuator -(2)EcoRI+SpeI
3
4

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

Nameconcentration[µg/mL]260/280260/230
PT181-attenuator EcoI+SpeI9.21.150.12
LaneDNAEnzyme
11kbpladder--
2pSB1C3EcoI+SpeI
3
4

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

Nameconcentration[µg/mL]260/280260/230
pSB1C310.01.840.40
LaneDNAEnzyme
11kbpladder--
2pSB4K5XbaI+PstI
3
4

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

Nameconcentration[µg/mL]260/280260/230
pSB4K57.61.810.32
LaneDNAEnzyme
11kbpladder--
2Pcon-RBS-tetR-DTXbaI+PstI
3
4

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

Nameconcentration[µg/mL]260/280260/230
Pcon-RBS-tetR-DT8.02.401.14
LaneDNAEnzyme
11kbpladder--
2pSB1C3EcoRI+PstI
3
4

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

Nameconcentration[µg/mL]260/280260/230
pSB1C39.12.170.35

Mutation PCR

Kinose

pC KaiABC(100pg)Prime STAR(2x)Primer(KaiA-Bsai MuT-Fwd)Primer(KaiA-Bsai MuT-Rev)MilliQtotal
1.025.01.01.022.050.0
1.525.01.01.021.550.0
2.025.01.01.021.050.0
PreDenatureDenatureAnnealingExtensioncycle
96°C98°C65°C72°C--
5m10s15s40s30cycles
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