Template:Kyoto/Notebook/Sep 16

From 2013.igem.org

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|experiment||pC181 attenuator(XbaI+PstI)||8.2µg/mL||pSB1C3(XbaI+PstI)||26.0µg/mL||xx µg/mL
|experiment||pC181 attenuator(XbaI+PstI)||8.2µg/mL||pSB1C3(XbaI+PstI)||26.0µg/mL||xx µg/mL
 +
|}
 +
===PCR===
 +
<!-- ここから -->
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!9/14 aptamer12-P(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|1.5||0.5||1.75||0.5||5.75||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/14 aptamer12-1M(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|1.7||0.5||1.75||0.5||6.05||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/14 Fusion1 attenuator(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|2.3||0.5||1.75||0.5||5.45||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/14 Fusion1 antisense(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|1.3||0.5||1.75||0.5||6.45||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/13 Fusion3m2 attenuator(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|1.7||0.5||1.75||0.5||6.05||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/13 Fusion6 antisense(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|1.6||0.5||1.75||0.5||6.15||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/12 plux(VR)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|2.4||0.5||1.75||0.5||5.35||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/9 Ptet antisense(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|2.2||0.5||1.75||0.5||5.55||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/12 Pcon-pT181 attenuator-DT(VF)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|1.4||0.5||1.75||0.5||6.35||10.5
 +
|}
 +
{| class="wikitable"
 +
!9/12 Pcon-pT181 attenuator-DT(VR)||Big Dye||5×Buffer||primer||MilliQ||total
 +
|-
 +
|1.4||0.5||1.75||0.5||6.35||10.5
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|96&deg;C||96&deg;C||50&deg;C||60&deg;C||--
 +
|-
 +
|2min||10s||5s||4min||30cycles
|}
|}
===Miniprep===
===Miniprep===

Revision as of 01:59, 26 September 2013

Contents

Sep 16

Liquid Culture

No name

Samplemedium
pSB1C3+P-KaiBC①Plusgrow medium(+CP)
pSB1C3+P-KaiBC②Plusgrow medium(+CP)

Colony PCR

No name

Samplebase pair
pSB1C3+P-KaiBC①283
pSB1C3+P-KaiBC②283
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
2min30s30s18s30cycles

Gel Extraction

No name

LaneDNAEnzyme
11kb ladder
2pT181 attenuatorXbaI+PstI
3pT181 attenuatorXbaI+PstI
4pT181 attenuatorXbaI+PstI
6pSB1C3EcoRI+SpeI
7pSB1C3EcoRI+SpeI
8pSB1C3EcoRI+SpeI
10pSB4K5XbaI+PstI
11pSB4K5XbaI+PstI
12pSB4K5XbaI+PstI

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

Nameconcentration[µg/mL]260/280260/230
pT181 attenuator(XbaI+PstI)8.21.620.11
pSB1C3(EcoRI+SpeI)49.51.480.18
pSB4K5(XbaI+PstI)18.11.730.57
DT(SpeI+PstI)70.41.361.02
aptamer12-1R-DT(SpeI+PstI)42.01.820.47
spinach-DT(SpeI+PstI)36.81.790.61

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
Dx-lux1136.71.461.28
RBS-laczα-DT237.61.881.70
pSB4K5148.41.551.09
Pcon-KBS-tetR-DT126.61.731.36
9/11Plac1160.01.781.28
9/15Plac2144.01.731.81
9/14Plac332.51.512.33
Ptet-RBS-laczα-DT231.31.841.57

PCR

No name

DNA9/6pT181 anti psB1C3(1)KOD plus10x bufferdNTPMgSO4GG1_1C3_fwd primerGG2_1C3_rev primerMilliQtotal
400pg10.52.52.51.50.750.7515.525
800pg20.52.52.51.50.750.7514.525
1000pg2.50.52.52.51.50.750.751425
nega00.52.52.51.50.750.7516.525
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2min10s30s30s30cycles
DNA9/6 12-1R aptamer-DTKOD plus10x bufferdNTPMgSO4GG1_1C3_fwd primerGG1_1C3_rev primerMilliQtotal
400pg10.52.52.51.50.750.7515.525
800pg20.52.52.51.50.750.7514.525
1000pg2.50.52.52.51.50.750.751425
NC00.52.52.51.50.750.7516.525
PreDenatureDenatureAnnealingExtensioncycle
94°C98°C57°C68°C--
2min10s30s30s30cycles

Electrophoresis

Yoshida

LaneSample
1100bp ladder
2antisense 1000pg
3antisense 800pg
4antisense 400pg
5tet aptamer-DT 1000pg
6tet aptamer-DT 800pg
7tet aptamer-DT 400pg
8NC

File:Igku xxxxxx.xxx

Restriction Enzyme Digestion

Tatsui

9/16 Pcon-RBS-tetR-DT(µL)EcoRI(µL)SpeI(µL)XbaI(µL)PstI(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts15.80011336.230
NC0.80000117.210
9/16 pSB4K5(µL)EcoRI(µL)SpeI(µL)XbaI(µL)PstI(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts13.50011338.530
NC0.70000117.310
8/22 pSB1C3(µL)EcoRI(µL)SpeI(µL)XbaI(µL)PstI(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts18.51001333.530
NC0.90000117.110
8/22 pSB1C3(µL)EcoRI(µL)SpeI(µL)XbaI(µL)PstI(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts18.51100333.530
NC0.90000117.110
8/21 pT181 attenuator2(µL)EcoRI(µL)SpeI(µL)XbaI(µL)PstI(µL)Buffer(µL)BSA(µL)MilliQ(µL)total(µL)
2 cuts7.411003314.630
NC0.40000117.610

Ligation

Tatsui

stateVectorInserterLigation High ver.2
experimentPcon-pT181anti-spinach-DT(XbaI+PstI)5.0µg/mLpSB4K5(XbaI+PstI)18.1µg/mLxx µg/mL
experimentpC181 attenuator(XbaI+PstI)8.2µg/mLpSB1C3(XbaI+PstI)26.0µg/mLxx µg/mL

PCR

No name

9/14 aptamer12-P(VF)Big Dye5×BufferprimerMilliQtotal
1.50.51.750.55.7510.5
9/14 aptamer12-1M(VF)Big Dye5×BufferprimerMilliQtotal
1.70.51.750.56.0510.5
9/14 Fusion1 attenuator(VF)Big Dye5×BufferprimerMilliQtotal
2.30.51.750.55.4510.5
9/14 Fusion1 antisense(VF)Big Dye5×BufferprimerMilliQtotal
1.30.51.750.56.4510.5
9/13 Fusion3m2 attenuator(VF)Big Dye5×BufferprimerMilliQtotal
1.70.51.750.56.0510.5
9/13 Fusion6 antisense(VF)Big Dye5×BufferprimerMilliQtotal
1.60.51.750.56.1510.5
9/12 plux(VR)Big Dye5×BufferprimerMilliQtotal
2.40.51.750.55.3510.5
9/9 Ptet antisense(VF)Big Dye5×BufferprimerMilliQtotal
2.20.51.750.55.5510.5
9/12 Pcon-pT181 attenuator-DT(VF)Big Dye5×BufferprimerMilliQtotal
1.40.51.750.56.3510.5
9/12 Pcon-pT181 attenuator-DT(VR)Big Dye5×BufferprimerMilliQtotal
1.40.51.750.56.3510.5
PreDenatureDenatureAnnealingExtensioncycle
96°C96°C50°C60°C--
2min10s5s4min30cycles

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
pSBiC3111.71.791.36
spinach-DT113.91.571.38
DT77.02.051.92
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