Template:Kyoto/Notebook/Sep 23

From 2013.igem.org

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<span class="author">じっけんしゃ</span>
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<span class="author">Hirano</span>
{| class="wikitable"
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!Lane||Sample||Enzyme1||Enzyme2
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[[File:igku_xxxxxx.xxx]]<br>
[[File:igku_xxxxxx.xxx]]<br>
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===Gel Extraction===
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<div class="experiment">
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<span class="author">Hirano</span>
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{| class="wikitable"
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!Name||concentration[&micro;g/mL]||260/280||260/230
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|-
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|Pcon atte-DT(E-1A)||166.7 ||1.51 ||0.99
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|-
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|Pcon atte-DT(2-1A)||122.1 ||1.75 ||1.20
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|-
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|Pcon anti spi DT(1-2A)||99.1 ||1.86 ||1.81
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|}
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Revision as of 04:55, 24 September 2013

Contents

Sep 23

Gel Extraction

No Name

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

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
9/22 Plac-DT181 atte-1204.0 2.00 1.95
9/22 Plac-DT181 atte-2195.3 1.84 1.62
9/22 Plac-DT181 anti-1188.2 1.87 2.15
9/22 Plac-DT181 anti-2165.0 1.95 2.66

Electrophoresis

Hirano

LaneSampleEnzyme1Enzyme2
1,6100bp ladder(Promega)
2Pcon-atte-DT (E-1A)
3Pcon-atte-DT (2-1A)
4Pcon-anti-spi-DT (1-2A)
5Pcon-anti-spi-DT (E-1A)

File:Igku xxxxxx.xxx

Gel Extraction

Hirano

Nameconcentration[µg/mL]260/280260/230
Pcon atte-DT(E-1A)166.7 1.51 0.99
Pcon atte-DT(2-1A)122.1 1.75 1.20
Pcon anti spi DT(1-2A)99.1 1.86 1.81