Template:Kyoto/Notebook/Sep 5

From 2013.igem.org

(Difference between revisions)
(→Sep 5)
(→Sep 5)
Line 251: Line 251:
|-
|-
|9/11 aptamer12-1M-2||Plusgrow medium(+CP)
|9/11 aptamer12-1M-2||Plusgrow medium(+CP)
 +
|}
 +
</div>
 +
 +
===Transformation===
 +
<div class="experiment">
 +
<span class="author">じっけんしゃ</span>
 +
{| class="wikitable"
 +
!Name||Sample(&micro;L)||Competent Cells(&micro;L)||Total(&micro;L)||Plate
 +
|-
 +
|9/8 Pcont+aptamer 12-1R-DT||1||10||11||Amp
 +
|-
 +
|9/8 Pcon+RBS-lacZα-DT||1||10||11||Amp
 +
|-
 +
|9/8 Pcon-pT181 attenuater+aptamer 12-1R-DT||1||10||11||Amp
 +
|-
 +
|9/8 Plac+RBS-lysis1-DT||2||20||22||CP
 +
|-
 +
|9/8 Plac+RBS-lysis2-DT||2||20||22||CP
 +
|-
 +
|9/8 Plac+aptamer 12-1R-DT||2||20||22||CP
 +
|-
 +
|9/8 Plac+spinach||2||20||22||CP
 +
|-
 +
|9/8 Plac+pT181 attenuater||2||20||22||CP
 +
|-
 +
|9/8 Plux+RBS-lysis1-DT||1||10||11||CP
 +
|-
 +
|9/8 Plux+RBS-lysis2-DT||1||10||11||CP
 +
|-
 +
|9/8 Ptet+RBS-lacZα-DT||1||10||11||CP
 +
|-
 +
|9/8 Ptet-pT181 anti+spinach-DT||1||10||11||CP
 +
|-
 +
|9/8 Pcon-pT181 attenuater+DT||1||10||11||CP
 +
|}
 +
 +
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">じっけんしゃ</span>
 +
{| class="wikitable"
 +
! ||9/8 Pcon-pT181 anti||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||8.2&micro;L|| 0&micro;L|| 1&micro;L|| 0&micro;L|| 1&micro;L|| 3&micro;L|| 3&micro;L|| 13.8&micro;L|| 30&micro;L
 +
|-
 +
|NC|| 0.4&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 1&micro;L|| 7.6&micro;L|| 10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||9/8 Pcon-spin-DT||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||7.9&micro;L|| 1&micro;L|| 1&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 3&micro;L||14.1&micro;L|| 30&micro;L
 +
|-
 +
|NC|| 0.4&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 1&micro;L||7.6&micro;L|| 10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 F1 atte||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts|| 6.5&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 1&micro;L|| 3&micro;L|| 3&micro;L||15.5&micro;L||30&micro;L
 +
|-
 +
|NC|| 0.3&micro;L||0&micro;L||0&micro;L|| 0&micro;L|| 0&micro;L||1&micro;L||1&micro;L||7.7&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 F3m2 atte||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||6.0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||16&micro;L||30&micro;L
 +
|-
 +
|NC|| 0.3&micro;L||0&micro;L||0&micro;L|| 0&micro;L|| 0&micro;L||1&micro;L||1&micro;L||7.7&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 F1 anti||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||6.7&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||15.3&micro;L||30&micro;L
 +
|-
 +
|NC|| 0.3&micro;L||0&micro;L||0&micro;L|| 0&micro;L|| 0&micro;L||1&micro;L||1&micro;L||7.7&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 F6 anti||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||6.0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||16&micro;L||30&micro;L
 +
|-
 +
|NC|| 0.3&micro;L||0&micro;L||0&micro;L|| 0&micro;L|| 0&micro;L||1&micro;L||1&micro;L||7.7&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 tetR aptamer 12_1M||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||5.3&micro;L||1&micro;L||1&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||16.7&micro;L||30&micro;L
 +
|-
 +
|NC|| 0.3&micro;L||0&micro;L||0&micro;L|| 0&micro;L|| 0&micro;L||1&micro;L||1&micro;L||7.7&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 tetR aptamer 12_P||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||13.3&micro;L||1&micro;L||1&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||8.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.7&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.3&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||9/4 spin-DT||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
||2 cuts||5.4&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||16.6&micro;L||30&micro;L
 +
|-
 +
|NC||0.3&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.7&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||9/7 RBS-lysis1-DT||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts|||10&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||12&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.5&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||9/6 RBS-lysis2-DT||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
||2 cuts||13&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||9&micro;L||30&micro;L
 +
|-
 +
|NC||0.7&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.3&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||9/7 RBS-lysis3-DT||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts|||10&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||12&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.5&micro;L||10&micro;L
 +
 +
|}
 +
{| class="wikitable"
 +
! ||8/20 J23100 RBS-lacZα-DT-(1)||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts|||7&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||16&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.5&micro;L||10&micro;L
 +
|-
 +
|}
 +
{| class="wikitable"
 +
! ||8/20 RBS-lacZα-DT||E||S||X||P||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||10&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||13&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.5&micro;L||10&micro;L
 +
|-
 +
|}
 +
 +
===5*M9(EDTA)===
 +
 +
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||
 +
|-
 +
|2||RBS-lysis1-DT||XbaI & PstI
 +
|-
 +
|3||RBS-lysis1-DT||XbaI & PstI
 +
|-
 +
|5||RBS-lysis3-DT||XbaI & PstI
 +
|-
 +
|6||RBS-lysis3-DT||XbaI & PstI
 +
|}
 +
[[File:igku_xxbeforexx.xxx]]<br>
 +
[[File:igku_xxafterxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|RBS-lysis1-DT (XbaI & PstI)||5.3||1.98||0.06
 +
|-
 +
|RBS-lysis3-DT (XbaI & PstI)||5.3||1.98||0.06
 +
|}
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||
 +
|-
 +
|3||Pcon-pT181 attenuator||EcoRI & SpeI
 +
|-
 +
|4||Pcon-pT181 attenuator||EcoRI & SpeI
 +
|-
 +
|6||Spinach-DT||XbaI & PstI
 +
|-
 +
|7||Spinach-DT||XbaI & PstI
 +
|-
 +
|9||RBS-lacZ&alpha;-DT||XbaI & PstI
 +
|-
 +
|10||RBS-lacZ&alpha;-DT||XbaI & PstI
 +
|}
 +
[[File:igku_xxbeforexx.xxx]]<br>
 +
[[File:igku_xxafterxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|Pcon-pT181 attenuator (EcoRI & SpeI)||5.2||1.82||0.36
 +
|-
 +
|Spinach-DT (XbaI & PstI)||8.0||1.87||0.25
 +
|-
 +
|RBS-lacZ&alpha;-DT (XbaI & PstI)||10.3||1.71||0.54
 +
|}
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||1kbp ladder||
 +
|-
 +
|2||Ptet-pT181 antisense||EcoRI & PstI
 +
|-
 +
|3||Ptet-pT181 antisense||EcoRI & PstI
 +
|-
 +
|5||pSB4K5||EcoRI & SpeI
 +
|-
 +
|6||pSB4K5||EcoRI & SpeI
 +
|}
 +
[[File:igku_xxbeforexx.xxx]]<br>
 +
[[File:igku_xxafterxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|Ptet-pT181 antisense(EcoRI & PstI)||30.6||1.84||1.16
 +
|-
 +
|pSB4K5 (EcoRI & SpeI)||21.8||1.87||0.98
 +
|}
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||1kbp ladder||
 +
|-
 +
|2||Pcon-pT181 attenuator||SpeI & PstI
 +
|-
 +
|3||Pcon-pT181 attenuator||SpeI & PstI
 +
|-
 +
|5||Plac||SpeI & PstI
 +
|-
 +
|6||Plac||SpeI & PstI
 +
|-
 +
|8||pSB1C3||EcoRI & SpeI
 +
|-
 +
|9||pSB1C3||EcoRI & SpeI
 +
|-
 +
|11||pSB1C3||XbaI & PstI
 +
|-
 +
|12||pSB1C3||XbaI & PstI
 +
|}
 +
[[File:igku_xxbeforexx.xxx]]<br>
 +
[[File:igku_xxafterxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|Pcon-pT181 attenuator (SpeI & PstI)||14.5||1.91||1.09
 +
|-
 +
|Plac (SpeI & PstI)||11.6||1.74||1.19
 +
|-
 +
|pSB1C3 (EcoRI & SpeI)||5.7||1.74||0.34
 +
|-
 +
|pSB1C3 (XbaI & PstI)||5.6||2.00||0.37
 +
|}
 +
 +
===Miniprep===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{|class="wikitable"
 +
!DNA||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|Pcon-pT181 anti||243.0||1.90||1.81
 +
|-
 +
|Pcon-Spinach-DT||253.7||1.57||1.51
 +
|}
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|9/8 Plac-spinach-DT-(1)||659
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5m||30s||30s||40s||30cycles
 +
|}
 +
</div>
 +
<div class="experiment">
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|9/8 Plac-spinach-DT-(2)||659
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5m||30s||30s||40s||30cycles
 +
|}
 +
</div>
 +
<div class="experiment">
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|9/8 Pcon-pt181atte-DT-(1)||584
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5m||30s||30s||40s||30cycles
 +
|}
 +
</div>
 +
 +
===Electrophoresis===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||--||--
 +
|-
 +
|2||8/19 lysis1||XbaI||PstI
 +
|-
 +
|3||8/19 lysis1||XbaI||--
 +
|-
 +
|4||8/19 lysis1||--||PstI
 +
|-
 +
|5||8/19 lysis1||--||--
 +
|-
 +
|6||8/19 lysis1(Gel Extraction Product)||XbaI||PstI
 +
|}
 +
</div>
 +
 +
===Liquid Culture===
 +
<div class="experiment">
 +
<span class="author">Hirano</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|JM109-1(Master plate)||LB medium
 +
|-
|}
|}
</div>
</div>

Revision as of 07:37, 24 September 2013

Contents

Sep 5

Colony PCR

No name

Samplebase pair
9/8 Plac-spinach-DT-(1)659
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles
Samplebase pair
9/8 Plac-spinach-DT-(2)659
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles
Samplebase pair
9/8 Pcon-pt181atte-DT-(1)584
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder----
28/19 lysis1XbaIPstI
38/19 lysis1XbaI--
48/19 lysis1--PstI
58/19 lysis1----
68/19 lysis1(Gel Extraction Product)XbaIPstI

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
Pcon-pT181 anti243.01.901.81
Pcon-Spinach-DT253.71.571.51

Ligation

Kojima

stateVectorInserterLigation High ver.2
experimentPconst13.2spinach-DT2.13.5
experimentPlac15.3spinach-DT2.13.5
experimentPconst13.2pT181 antisense (XbaI & PstI)6.63.5
experimentPlac15.3pT181 antisense (XbaI & PstI)6.63.5
experimentPtet30.1pT181 antisense (XbaI & PstI)6.63.5
experimentPconst13.2pT181 attenuator11.73.5
experimentPlac15.3pT181 attenuator11.73.5
experimentPtet30.1RBS-lacZα-DT3.03.5

incubate 16 °C 1 hour

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder
2RBS-lysis1-DTXbaI & PstI
3RBS-lysis1-DTXbaI & PstI
5RBS-lysis3-DTXbaI & PstI
6RBS-lysis3-DTXbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
RBS-lysis1-DT (XbaI & PstI)5.31.980.06
RBS-lysis3-DT (XbaI & PstI)5.31.980.06
LaneDNAEnzyme
1100bp ladder
3Pcon-pT181 attenuatorEcoRI & SpeI
4Pcon-pT181 attenuatorEcoRI & SpeI
6Spinach-DTXbaI & PstI
7Spinach-DTXbaI & PstI
9RBS-lacZα-DTXbaI & PstI
10RBS-lacZα-DTXbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
Pcon-pT181 attenuator (EcoRI & SpeI)5.21.820.36
Spinach-DT (XbaI & PstI)8.01.870.25
RBS-lacZα-DT (XbaI & PstI)10.31.710.54
LaneDNAEnzyme
11kbp ladder
2Ptet-pT181 antisenseEcoRI & PstI
3Ptet-pT181 antisenseEcoRI & PstI
5pSB4K5EcoRI & SpeI
6pSB4K5EcoRI & SpeI

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

Nameconcentration[µg/mL]260/280260/230
Ptet-pT181 antisense(EcoRI & PstI)30.61.841.16
pSB4K5 (EcoRI & SpeI)21.81.870.98
LaneDNAEnzyme
11kbp ladder
2Pcon-pT181 attenuatorSpeI & PstI
3Pcon-pT181 attenuatorSpeI & PstI
5PlacSpeI & PstI
6PlacSpeI & PstI
8pSB1C3EcoRI & SpeI
9pSB1C3EcoRI & SpeI
11pSB1C3XbaI & PstI
12pSB1C3XbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
Pcon-pT181 attenuator (SpeI & PstI)14.51.911.09
Plac (SpeI & PstI)11.61.741.19
pSB1C3 (EcoRI & SpeI)5.71.740.34
pSB1C3 (XbaI & PstI)5.62.000.37

Colony PCR

Hirano

Samplebase pair
8/9 BBa_J23100 3--
8/9 BBa_J23100 4--
8/9 BBa_J23100 5--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

Liquid Culture

Nakamoto

Samplemedium
9/11Fusion6 antisense-1Plusgrow medium(+CP)
Plac(BBa-R0011)-3Plusgrow medium(+Amp)
Plac(BBa-R0011)-4Plusgrow medium(+Amp)
Fusion3m2 attenuator -1Plusgrow medium(+CP)　
Pλ-luxI-2Plusgrow medium(+Amp)　
9/11 aptamer12-1M-2Plusgrow medium(+CP)

Transformation

じっけんしゃ

NameSample(µL)Competent Cells(µL)Total(µL)Plate
9/8 Pcont+aptamer 12-1R-DT11011Amp
9/8 Pcon+RBS-lacZα-DT11011Amp
9/8 Pcon-pT181 attenuater+aptamer 12-1R-DT11011Amp
9/8 Plac+RBS-lysis1-DT22022CP
9/8 Plac+RBS-lysis2-DT22022CP
9/8 Plac+aptamer 12-1R-DT22022CP
9/8 Plac+spinach22022CP
9/8 Plac+pT181 attenuater22022CP
9/8 Plux+RBS-lysis1-DT11011CP
9/8 Plux+RBS-lysis2-DT11011CP
9/8 Ptet+RBS-lacZα-DT11011CP
9/8 Ptet-pT181 anti+spinach-DT11011CP
9/8 Pcon-pT181 attenuater+DT11011CP

Restriction Enzyme Digestion

じっけんしゃ

9/8 Pcon-pT181 antiESXPBSABufferMilliQtotal
2 cuts8.2µL 0µL 1µL 0µL 1µL 3µL 3µL 13.8µL 30µL
NC 0.4µL 0µL 0µL 0µL 0µL 1µL 1µL 7.6µL 10µL
9/8 Pcon-spin-DTESXPBSABufferMilliQtotal
2 cuts7.9µL 1µL 1µL 0µL 0µL 3µL 3µL14.1µL 30µL
NC 0.4µL 0µL 0µL 0µL 0µL 1µL 1µL7.6µL 10µL
8/21 F1 atteESXPBSABufferMilliQtotal
2 cuts 6.5µL 0µL 0µL 1µL 1µL 3µL 3µL15.5µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 F3m2 atteESXPBSABufferMilliQtotal
2 cuts6.0µL0µL0µL1µL1µL3µL3µL16µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 F1 antiESXPBSABufferMilliQtotal
2 cuts6.7µL0µL0µL1µL1µL3µL3µL15.3µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 F6 antiESXPBSABufferMilliQtotal
2 cuts6.0µL0µL0µL1µL1µL3µL3µL16µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 tetR aptamer 12_1MESXPBSABufferMilliQtotal
2 cuts5.3µL1µL1µL0µL0µL3µL3µL16.7µL30µL
NC 0.3µL0µL0µL 0µL 0µL1µL1µL7.7µL10µL
8/21 tetR aptamer 12_PESXPBSABufferMilliQtotal
2 cuts13.3µL1µL1µL0µL0µL3µL3µL8.7µL30µL
NC0.7µL0µL0µL0µL0µL1µL1µL7.3µL10µL
9/4 spin-DTESXPBSABufferMilliQtotal
2 cuts5.4µL0µL0µL1µL1µL3µL3µL16.6µL30µL
NC0.3µL0µL0µL0µL0µL1µL1µL7.7µL10µL
9/7 RBS-lysis1-DTESXPBSABufferMilliQtotal
2 cuts10µL0µL0µL1µL1µL3µL3µL12µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL
9/6 RBS-lysis2-DTESXPBSABufferMilliQtotal
2 cuts13µL0µL0µL1µL1µL3µL3µL9µL30µL
NC0.7µL0µL0µL0µL0µL1µL1µL7.3µL10µL
9/7 RBS-lysis3-DTESXPBSABufferMilliQtotal
2 cuts10µL0µL0µL1µL1µL3µL3µL12µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL
8/20 J23100 RBS-lacZα-DT-(1)ESXPBSABufferMilliQtotal
2 cuts7µL0.5µL0.5µL0µL0µL3µL3µL16µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL
8/20 RBS-lacZα-DTESXPBSABufferMilliQtotal
2 cuts10µL0.5µL0.5µL0µL0µL3µL3µL13µL30µL
NC0.5µL0µL0µL0µL0µL1µL1µL7.5µL10µL

5*M9(EDTA)

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder
2RBS-lysis1-DTXbaI & PstI
3RBS-lysis1-DTXbaI & PstI
5RBS-lysis3-DTXbaI & PstI
6RBS-lysis3-DTXbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
RBS-lysis1-DT (XbaI & PstI)5.31.980.06
RBS-lysis3-DT (XbaI & PstI)5.31.980.06
LaneDNAEnzyme
1100bp ladder
3Pcon-pT181 attenuatorEcoRI & SpeI
4Pcon-pT181 attenuatorEcoRI & SpeI
6Spinach-DTXbaI & PstI
7Spinach-DTXbaI & PstI
9RBS-lacZα-DTXbaI & PstI
10RBS-lacZα-DTXbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
Pcon-pT181 attenuator (EcoRI & SpeI)5.21.820.36
Spinach-DT (XbaI & PstI)8.01.870.25
RBS-lacZα-DT (XbaI & PstI)10.31.710.54
LaneDNAEnzyme
11kbp ladder
2Ptet-pT181 antisenseEcoRI & PstI
3Ptet-pT181 antisenseEcoRI & PstI
5pSB4K5EcoRI & SpeI
6pSB4K5EcoRI & SpeI

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

Nameconcentration[µg/mL]260/280260/230
Ptet-pT181 antisense(EcoRI & PstI)30.61.841.16
pSB4K5 (EcoRI & SpeI)21.81.870.98
LaneDNAEnzyme
11kbp ladder
2Pcon-pT181 attenuatorSpeI & PstI
3Pcon-pT181 attenuatorSpeI & PstI
5PlacSpeI & PstI
6PlacSpeI & PstI
8pSB1C3EcoRI & SpeI
9pSB1C3EcoRI & SpeI
11pSB1C3XbaI & PstI
12pSB1C3XbaI & PstI

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

Nameconcentration[µg/mL]260/280260/230
Pcon-pT181 attenuator (SpeI & PstI)14.51.911.09
Plac (SpeI & PstI)11.61.741.19
pSB1C3 (EcoRI & SpeI)5.71.740.34
pSB1C3 (XbaI & PstI)5.62.000.37

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
Pcon-pT181 anti243.01.901.81
Pcon-Spinach-DT253.71.571.51

Colony PCR

No name

Samplebase pair
9/8 Plac-spinach-DT-(1)659
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles
Samplebase pair
9/8 Plac-spinach-DT-(2)659
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles
Samplebase pair
9/8 Pcon-pt181atte-DT-(1)584
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5m30s30s40s30cycles

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder----
28/19 lysis1XbaIPstI
38/19 lysis1XbaI--
48/19 lysis1--PstI
58/19 lysis1----
68/19 lysis1(Gel Extraction Product)XbaIPstI

Liquid Culture

Hirano

Samplemedium
JM109-1(Master plate)LB medium
Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Sep_5"