Template:Kyoto/Notebook/Sep 17

From 2013.igem.org

(Difference between revisions)
(→PCR)
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!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
|-
|-
-
|experiment||8/28 Plux(S+P)||45µg/mL||9/13 RBS-lysis1-DT (X+P)||7.8µg/mL||xx µg/mL
+
|experiment||8/28 Plux(SpeI+PstI)||45µg/mL||9/13 RBS-lysis1-DT (XbaI+PstI)||7.8µg/mL||xx µg/mL
|-
|-
-
|experiment||8/28 Plux(S+P)||45µg/mL||9/13 RBS-lysis2-DT (X+P)||11.8µg/mL||xx µg/mL
+
|experiment||8/28 Plux(SpeI+PstI)||45µg/mL||9/13 RBS-lysis2-DT (XbaI+PstI)||11.8µg/mL||xx µg/mL
|-
|-
-
|experiment||8/28 Plux(S+P)||45µg/mL||9/13 RBS-lysis3-DT (X+P)||11.8µg/mL||xx µg/mL
+
|experiment||8/28 Plux(SpeI+PstI)||45µg/mL||9/13 RBS-lysis3-DT (XbaI+PstI)||11.8µg/mL||xx µg/mL
|-
|-
-
|experiment||9/14 Plac(S+P)||21µg/mL||9/16 PT181 atte-DT (X+P)||8.2µg/mL||xx µg/mL
+
|experiment||9/14 Plac(SpeI+PstI)||21µg/mL||9/16 PT181 atte-DT (XbaI+PstI)||8.2µg/mL||xx µg/mL
|-
|-
-
|experiment||9/14 Plac(S+P)||21µg/mL||9/3 PT181 anti (X+P)||6.6µg/mL||xx µg/mL
+
|experiment||9/14 Plac(SpeI+PstI)||21µg/mL||9/3 PT181 anti (XbaI+PstI)||6.6µg/mL||xx µg/mL
|-
|-
-
|experiment||9/14 Plac(S+P)||21µg/mL||9/10 apt12-1R-DT(X+P)||15.6µg/mL||xx µg/mL
+
|experiment||9/14 Plac(SpeI+PstI)||21µg/mL||9/10 apt12-1R-DT(XbaI+PstI)||15.6µg/mL||xx µg/mL
|-
|-
-
|experiment||9/13 pSB1C3(X+P)||26.0µg/mL||9/15 Pcon-PT181 anti-spin-DT(X+P)||5.0µg/mL||xx µg/mL
+
|experiment||9/13 pSB1C3(XbaI+PstI)||26.0µg/mL||9/15 Pcon-PT181 anti-spin-DT(XbaI+PstI)||5.0µg/mL||xx µg/mL
|-
|-
-
|experiment||9/16 pSB1C3(E+S)||49.5µg/mL||9/15 Pcon-spin-DT(E+S)||9.0µg/mL||xx µg/mL
+
|experiment||9/16 pSB1C3(EcoRI+SpeI)||49.5µg/mL||9/15 Pcon-spin-DT(EcoRI+SpeI)||9.0µg/mL||xx µg/mL
|-
|-
-
|experiment||9/16 pSB1C3(E+S)||49.5µg/mL||9/15 Pcon-RBS-tetR-DT(E+S)||59.0µg/mL||xx µg/mL
+
|experiment||9/16 pSB1C3(EcoRI+SpeI)||49.5µg/mL||9/15 Pcon-RBS-tetR-DT(EcoRI+SpeI)||59.0µg/mL||xx µg/mL
|-
|-
-
|experiment||9/16 pSB1C3(E+S)||49.5µg/mL||9/15 Pcon-PT181-atte-DT(E+S)||21.0µg/mL||xx µg/mL
+
|experiment||9/16 pSB1C3(EcoRI+SpeI)||49.5µg/mL||9/15 Pcon-PT181-atte-DT(EcoRI+SpeI)||21.0µg/mL||xx µg/mL
|-
|-
-
|experiment||9/16 pSB1C3(E+S)||49.5µg/mL||9/15 Pcon-apt12-112-DT(E+S)||17.0µg/mL||xx µg/mL
+
|experiment||9/16 pSB1C3(EcoRI+SpeI)||49.5µg/mL||9/15 Pcon-apt12-112-DT(EcoRI+SpeI)||17.0µg/mL||xx µg/mL
|}
|}
</div>
</div>

Revision as of 06:35, 24 September 2013

Contents

Sep 17

Liquid Culture

Tatsui and Hirano

Samplemedium
Spi-DTLB medium(+CP)
Pcon-Spi-DT,Pcon-pT181-anti-spi-DTLB medium(+Amp)　

PCR

Hirano

Pcon-tet ap-DT(VF)BigDye5x bufferprimer(VF)MilliQtotal
1.30.51.750.50.4510.5
PreDenatureDenatureAnnealingExtensioncycle
96°C96°C50°C60°C--
2m10s5s4m30cycles
Pcon-tet ap-DT(VR)BigDye5x bufferprimer(VR)MilliQtotal
1.30.51.750.50.4510.5
PreDenatureDenatureAnnealingExtensioncycle
96°C96°C50°C60°C--
2m10s5s4m30cycles

Electrophoresis

じっけんしゃ

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)45µg/mL9/13 RBS-lysis1-DT (XbaI+PstI)7.8µg/mLxx µg/mL
experiment8/28 Plux(SpeI+PstI)45µg/mL9/13 RBS-lysis2-DT (XbaI+PstI)11.8µg/mLxx µg/mL
experiment8/28 Plux(SpeI+PstI)45µg/mL9/13 RBS-lysis3-DT (XbaI+PstI)11.8µg/mLxx µg/mL
experiment9/14 Plac(SpeI+PstI)21µg/mL9/16 PT181 atte-DT (XbaI+PstI)8.2µg/mLxx µg/mL
experiment9/14 Plac(SpeI+PstI)21µg/mL9/3 PT181 anti (XbaI+PstI)6.6µg/mLxx µg/mL
experiment9/14 Plac(SpeI+PstI)21µg/mL9/10 apt12-1R-DT(XbaI+PstI)15.6µg/mLxx µg/mL
experiment9/13 pSB1C3(XbaI+PstI)26.0µg/mL9/15 Pcon-PT181 anti-spin-DT(XbaI+PstI)5.0µg/mLxx µg/mL
experiment9/16 pSB1C3(EcoRI+SpeI)49.5µg/mL9/15 Pcon-spin-DT(EcoRI+SpeI)9.0µg/mLxx µg/mL
experiment9/16 pSB1C3(EcoRI+SpeI)49.5µg/mL9/15 Pcon-RBS-tetR-DT(EcoRI+SpeI)59.0µg/mLxx µg/mL
experiment9/16 pSB1C3(EcoRI+SpeI)49.5µg/mL9/15 Pcon-PT181-atte-DT(EcoRI+SpeI)21.0µg/mLxx µg/mL
experiment9/16 pSB1C3(EcoRI+SpeI)49.5µg/mL9/15 Pcon-apt12-112-DT(EcoRI+SpeI)17.0µg/mLxx µg/mL

Gel Extraction

No Name

LaneDNAEnzyme
11kbpladder--
2Pt181 atte -(2)E+S
3
4

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

Nameconcentration[µg/mL]260/280260/230
PT181-atte E+S9.21.150.12
LaneDNAEnzyme
11kbpladder--
2pSB1C3E+S
3
4

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

Nameconcentration[µg/mL]260/280260/230
pSB1C310.01.840.40
LaneDNAEnzyme
11kbpladder--
2pSB4K5X+P
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-DTX+P
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--
2pSB1C3E+P
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
PreDenatureDenatureAnnealingExtensioncycle
96°C98°C65°C72°C--
5m10s15s40s30cycles
pC KaiABC(150pg)Prime STAR(2x)Primer(KaiA-Bsai MuT-Fwd)Primer(KaiA-Bsai MuT-Rev)MilliQtotal
1.525.01.01.021.550.0
PreDenatureDenatureAnnealingExtensioncycle
96°C98°C65°C72°C--
5m10s15s40s30cycles

</div>

pC KaiABC(200pg)Prime STAR(2x)Primer(KaiA-Bsai MuT-Fwd)Primer(KaiA-Bsai MuT-Rev)MilliQtotal
225.01.01.021.050.0
PreDenatureDenatureAnnealingExtensioncycle
96°C98°C65°C72°C--
5m10s15s40s30cycles
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