Template:Kyoto/Notebook/Sep 23

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===Electrophoresis===
===Electrophoresis===
   
   
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<div class="experiment">
<div class="experiment">
<span class="author">Hirano</span>
<span class="author">Hirano</span>
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!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
|-
|-
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|100bp ladder|| || ||  
+
|1||100bp ladder|| ||  
|-
|-
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|9/8 Pcon-pT181 anti(E-1A)|| || ||
+
|2||9/8 Pcon-pT181 anti(E-1A)|| ||
|-
|-
-
|8/20 Ptet(1)(2-S)|| || ||
+
|3||8/20 Ptet(1)(2-S)|| ||
|-
|-
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|9/16 Plac2(E-1A)|| || ||
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|4||9/16 Plac2(E-1A)|| ||
|}
|}
[[File:igku_9231.png]]<br>
[[File:igku_9231.png]]<br>
</div>
</div>
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===Transformation===
===Transformation===

Revision as of 12:37, 26 September 2013

Contents

Sep 23

Transformation

No name

NameSampleCompetent CellsPlate
9/14 pSB1C3＋PkaiBC2 µL20 µL--
9/14 pSB1C3＋RpaB2 µL20 µL--
9/14 pkaiBC＋Rbs-GFP-DT2 µL20 µL--

Gel Extraction

No Name

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

Igku Sep23 Gel Extraction1.jpg
Igku Sep23 Gel Extraction2.jpg

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)

Igku 9232.png

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

Electrophoresis

Hirano

LaneSampleEnzyme1Enzyme2
1100bp ladder
29/8 Pcon-pT181 anti(E-1A)
38/20 Ptet(1)(2-S)
49/16 Plac2(E-1A)

Igku 9231.png

Transformation

No Name

NameSampleCompetent CellsTotalPlate
pSB1C32µL20µL22µL

Ligation

No Name

stateVectorInserterLigation High ver.2
experiment9/19 Plac S+P24.6 µg/mL9/18 ape12_1R-DT X+P4.5 µg/mL2 µg/mL
experiment9/21 RBS-GFP-DT E+X88.4 µg/mL9/15 Pcon-pT181 atte E+S21 µg/mL2 µg/mL
experiment9/13 PSB1C3 X+P26.0 µg/mL9/16 pT181 atte X+P8.2 µg/mL2 µg/mL


Colony PCR

Tatsui

Samplebase pair
9/22 Pcon-PT181 attenuator12-1R-DT(1~8)--
9/22 Pcon-PT181 attenuator(1~4)--
9/20 Plac＋aptamer12-1R-DT(1~4)--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s54s30cycle
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