Template:Kyoto/Notebook/Sep 23

From 2013.igem.org

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|experiment||9/13 DT E+X||25.4 µg/mL||9/22 Pcon-DT181 anti E+S||46.8 µg/mL||2.3 µg/mL
|experiment||9/13 DT E+X||25.4 µg/mL||9/22 Pcon-DT181 anti E+S||46.8 µg/mL||2.3 µg/mL
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|}
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</div>
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===PCR===
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<!-- ここから -->
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<div class="experiment">
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<span class="author">No name</span>
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{| class="wikitable"
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!genome DNA||KOD plus||10x buffer||dNTP||MgSO4||SasA_fwd primer||SasA_rev primer||MilliQ||total
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|-
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|Pcon-atte-DT (E-1A) ||0.5||2.5||2.5||1.5||0.75||0.75||77.5 ||25
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|-
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|Pcon-atte-DT (2-1A) ||0.5||2.5||2.5||1.5||0.75||0.75||77.5 ||25
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|-
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|Pcon-anti-spin-DT (1-2A) ||0.5||2.5||2.5||1.5||0.75||0.75||77.5 ||25
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|-
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|Pcon-anti-spin-DT (E-1A) ||0.5||2.5||2.5||1.5||0.75||0.75||77.5 ||25
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|}
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{| class="wikitable"
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!PreDenature||Denature||Annealing||Extension||cycle
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|-
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|94　&deg;C||98　&deg;C||57　&deg;C||68　&deg;C||--
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|-
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|2min　||10s　||30s　||45s　||30cycle
|}
|}
</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 04:30, 24 September 2013

Contents

Sep 23

Gel Extraction

じっけんしゃ

LaneDNAEnzyme
1 1kbp ladder
3-4 Pcon-RBS-tetR-PT

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

Nameconcentration[µg/mL]260/280260/230
Pcon-PT181 anti E+S46.7 1.06 0.93
Pcon-RBS-tetR-DT E+X119.3 0.99
RBS-8FP-DT E+X87.7 1.00 0.98
ap42_1R-DT E+X68.8 1.32 1.03
Pcon-RBS-tetR-DT 1-2 A203.0 1.66 1.79
Pcon-RBS-tetR-DT 3-2 A67.7 2.64 1.28
Pcon-RBS-GFP-DT 1-S A185.6 1.88 2.04
Pcon-RBS-GFP-DT 2-S A298.6 1.68 1.55
Pcon-tetR-DT E-2 A202.4 1.57 1.66

PCR

No name

genome DNAKOD plus10x bufferdNTPMgSO4SasA_fwd primerSasA_rev primerMilliQtotal
9/8 Pcon-pT181 anti 0.52.52.51.50.750.7515.5 25
8/20 Ptet(1) 0.52.52.51.50.750.7515.5 25
9/16 Plac 2 0.52.52.51.50.750.7515.5 25
PreDenatureDenatureAnnealingExtensioncycle
94　°C98　°C57　°C68　°C--
2min　10sec　30sec　20sec　30cycles

Ligation

Yoshida

stateVectorInserterLigation High ver.2
experiment9/21 Pcon-RBS-GFP-DT E+X47.5 µg/mL9/15 Pcon-RBS-tetR-DT E+S59 µg/mL2.3 µg/mL
experiment9/21 Pcon-RBS-GFP-DT E+X47.5 µg/mL9/15 Pcon-apt12_1R-DT E+S17 µg/mL2.4 µg/mL
experiment9/21 Pcon-RBS-GFRDT E+X2.1 µg/mL9/15 Pcon-DT181 atte-DT21 µg/mL3.0 µg/mL
experiment9/22 apt 12_1R-DT E+X67.8 µg/mL9/18 Pcon-DT181 atte E+S5.7 µg/mL3.5 µg/mL
experiment9/22RBS-GFP-DT E+X87.7 µg/mL9/18 Pcon-DT18 atte E+S6.7 µg/mL3.5 µg/mL
experiment9/13 DT E+X25.4 µg/mL9/22 Pcon-DT181 anti E+S46.8 µg/mL2.3 µg/mL

PCR

No name

genome DNAKOD plus10x bufferdNTPMgSO4SasA_fwd primerSasA_rev primerMilliQtotal
Pcon-atte-DT (E-1A) 0.52.52.51.50.750.7577.5 25
Pcon-atte-DT (2-1A) 0.52.52.51.50.750.7577.5 25
Pcon-anti-spin-DT (1-2A) 0.52.52.51.50.750.7577.5 25
Pcon-anti-spin-DT (E-1A) 0.52.52.51.50.750.7577.5 25
PreDenatureDenatureAnnealingExtensioncycle
94　°C98　°C57　°C68　°C--
2min　10s　30s　45s　30cycle
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