Template:Kyoto/Notebook/Sep 5

From 2013.igem.org

(Difference between revisions)
(→Sep 5)
(→Sep 5)
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==Sep 5==
==Sep 5==
 +
===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>
 +
 +
===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
 +
|}
 +
===Ligation===
===Ligation===
<div class="experiment">
<div class="experiment">
Line 24: Line 100:
|}
|}
incubate 16 &deg;C 1 hour  
incubate 16 &deg;C 1 hour  
 +
</div>
 +
 +
===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
 +
|}
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">Hirano</span>
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|8/9 BBa_J23100 3||--
 +
|-
 +
|8/9 BBa_J23100 4||--
 +
|-
 +
|8/9 BBa_J23100 5||--
 +
|-
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
</div>
 +
 +
===Liquid Culture===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|9/11Fusion6 antisense-1||Plusgrow medium(+CP)
 +
|-
 +
|Plac(BBa-R0011)-3||Plusgrow medium(+Amp)
 +
|-
 +
|Plac(BBa-R0011)-4||Plusgrow medium(+Amp)
 +
|-
 +
|Fusion3m2 attenuator -1||Plusgrow medium(+CP)　
 +
|-
 +
|P&lambda;-luxI-2||Plusgrow medium(+Amp)　
 +
|-
 +
|9/11 aptamer12-1M-2||Plusgrow medium(+CP)
 +
|}
</div>
</div>

Revision as of 07:25, 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)
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