Template:Kyoto/Notebook/Sep 17

From 2013.igem.org

(Difference between revisions)
(Gel Extraction)
(PCR)
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<div class="experiment">
<div class="experiment">
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<span class="author">No name</span>
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<span class="author">Kinose</span>
{| class="wikitable"
{| class="wikitable"
-
!genome DNA||KOD plus||10x buffer||dNTP||MgSO4||SasA_fwd primer||SasA_rev primer||MilliQ||total
+
!pC KaiABC(100pg)||Prime STAR(2x)||Primer(KaiA-Bsai MuT-Fwd)||Primer(KaiA-Bsai MuT-Rev)||MilliQ||total
|-
|-
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| ||0.5||2.5||2.5||1.5||0.75||0.75|| ||25
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|1.0||25.0||1.0||1.0||22.0||50.0
|}
|}
{| class="wikitable"
{| class="wikitable"
!PreDenature||Denature||Annealing||Extension||cycle
!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
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| &deg;C|| &deg;C|| &deg;C|| &deg;C||--
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|96&deg;C||98&deg;C||65&deg;C||72&deg;C||--
|-
|-
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|(◎Д◎) || || || ||
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|5m||10s||15s||40s||30cycles
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|}
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</div>
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<div class="experiment">
 +
{| class="wikitable"
 +
!pC KaiABC(150pg)||Prime STAR(2x)||Primer(KaiA-Bsai MuT-Fwd)||Primer(KaiA-Bsai MuT-Rev)||MilliQ||total
 +
|-
 +
|1.5||25.0||1.0||1.0||21.5||50.0
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|96&deg;C||98&deg;C||65&deg;C||72&deg;C||--
 +
|-
 +
|5m||10s||15s||40s||30cycles
 +
|}
 +
</div>
 +
<!-- ここまでをコピーしてね -->
 +
</div>
 +
<div class="experiment">
 +
{| class="wikitable"
 +
!pC KaiABC(200pg)||Prime STAR(2x)||Primer(KaiA-Bsai MuT-Fwd)||Primer(KaiA-Bsai MuT-Rev)||MilliQ||total
 +
|-
 +
|2||25.0||1.0||1.0||21.0||50.0
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|96&deg;C||98&deg;C||65&deg;C||72&deg;C||--
 +
|-
 +
|5m||10s||15s||40s||30cycles
|}
|}
</div>
</div>
<!-- ここまでをコピーしてね -->
<!-- ここまでをコピーしてね -->

Revision as of 02:39, 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

Ligation

No Name

stateVectorInserterLigation High ver.2
8/28 namexx µg/mLnamexx µg/mLxx µg/mL
NCnamexx µg/mLnamexx µg/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

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