Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Sep 6)
Line 1: Line 1:
==Sep 6==
==Sep 6==
 +
===Liquid Culture===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|8/21 RBS-Lysis2-9||Plusgrow medium(+Amp)
 +
|-
 +
|8/21 RBS-Lysis2-12||Plusgrow medium(+Amp)
 +
|-
 +
|8/21 RBS-Lysis2-14||Plusgrow medium(+Amp)　
 +
|}
 +
</div>
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/27 Pcon-pT181 antisense-(1)||542
 +
|-
 +
|8/26 pT181 attenuator-DT-(1)||738
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
[[File:igku_Aug19electrophoresis1.png]]
 +
</div>
 +
 +
===Miniprep===
 +
<div class="experiment">
 +
<span class="author">No Name</span>
 +
{|class="wikitable"
 +
!DNA||concentration [&micro;g/mL]||260/280||260/230
 +
|-
 +
|Plac||153.7||2.03||2.10
 +
|-
 +
|P&lambda;-luxⅠ-(1)||76.5||2.07||2.06
 +
|-
 +
|P&lambda;-luxⅠ-(2)||86.4||2.08||2.10
 +
|-
 +
|DT||166.2||1.65||2.12
 +
|-
 +
|J23100||190.1||1.68||2.34
 +
|}
 +
</div>
 +
===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
<div class="experiment">
<div class="experiment">
Line 29: Line 81:
|}
|}
*at 37&deg;C, for 1h
*at 37&deg;C, for 1h
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/27 Pcon-pT181 antisense-(1)||542
 +
|-
 +
|8/26 pT181 attenuator-DT-(1)||738
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
[[File:igku_Aug19electrophoresis1.png]]
 +
</div>
 +
 +
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|2||rowspan="2"|8/20 RBS-lysis1-(1)||rowspan="2"|EcoRI & SpeI
 +
|-
 +
|3
 +
|-
 +
|4||--||--
 +
|-
 +
|5||rowspan="2"|8/28 RBS-GFP-DT-(2)||rowspan="2"|XbaI & PstI
 +
|-
 +
|6
 +
|}
 +
[[File:igku_xxbeforexx.xxx]]<br>
 +
[[File:igku_xxafterxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|2||rowspan="2"|8/28 Plux||rowspan="2"|SpeI & PstI
 +
|-
 +
|3
 +
|-
 +
|4||--||--
 +
|-
 +
|5||rowspan="2"|8/20 RBS-tetR-DT-(1)||rowspan="2"|XbaI & PstI
 +
|-
 +
|6
 +
|}
 +
[[File:igku_xxbeforexx.xxx]]<br>
 +
[[File:igku_xxafterxx.xxx]]<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|Pcon-RBS-GFP-DT (EcoRI&SpeI)||7.1||1.88||0.07
 +
|-
 +
|Pcon-RBS-luxR-DT (EcoRI&XbaI)||15.3||1.93||0.70
 +
|-
 +
|Plux (SpeI&PstI)||4.5||1.79||0.31
 +
|-
 +
|RBS-GFP-DT (XbaI&PstI)||6.0||2.16||0.03
 +
|-
 +
|RBS-tetR-DT (XbaI&PstI)||7.6||1.91||0.38
 +
|}
 +
</div>
 +
 +
 +
===Liquid Culture===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|8/21 RBS-Lysis2-9||Plusgrow medium(+Amp)
 +
|-
 +
|8/21 RBS-Lysis2-12||Plusgrow medium(+Amp)
 +
|-
 +
|8/21 RBS-Lysis2-14||Plusgrow medium(+Amp)　
 +
|}
 +
</div>
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/27 Pcon-pT181 antisense-(1)||542
 +
|-
 +
|8/26 pT181 attenuator-DT-(1)||738
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
[[File:igku_Aug19electrophoresis1.png]]
 +
</div>
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/27 Pcon-pT181 antisense-(1)||542
 +
|-
 +
|8/26 pT181 attenuator-DT-(1)||738
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
[[File:igku_Aug19electrophoresis1.png]]
 +
</div>

Revision as of 09:26, 24 September 2013

Contents

Sep 6

Liquid Culture

No name

Samplemedium
8/21 RBS-Lysis2-9Plusgrow medium(+Amp)
8/21 RBS-Lysis2-12Plusgrow medium(+Amp)
8/21 RBS-Lysis2-14Plusgrow medium(+Amp)　

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

Miniprep

No Name

DNAconcentration [µg/mL]260/280260/230
Plac153.72.032.10
Pλ-luxⅠ-(1)76.52.072.06
Pλ-luxⅠ-(2)86.42.082.10
DT166.21.652.12
J23100190.11.682.34

Restriction Enzyme Digestion

Tatsui

9/6 RBS-lysis2-DT (78.3 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts22µL0µL0µL1µL1µL3µL3µL0µL30µL
NC1.3µL0µL0µL0µL0µL1µL1µL6.7µL10µL
  • at 37°C, for 1h
aptamer-DT (251.7 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts0.4µL0µL0µL1µL1µL3µL3µL14.1µL30µL
NC0.4µL0µL0µL0µL0µL1µL1µL7.6µL10µL
  • at 37°C, for 1h
Pλ-luxI (86.4 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts23.1µL1µL1µL0µL0µL3µL3µL1.9µL30µL
NC1.2µL0µL0µL0µL0µL1µL1µL6.8µL10µL
  • at 37°C, for 1h

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder--
28/20 RBS-lysis1-(1)EcoRI & SpeI
3
4----
58/28 RBS-GFP-DT-(2)XbaI & PstI
6

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

LaneDNAEnzyme
1100bp ladder--
28/28 PluxSpeI & PstI
3
4----
58/20 RBS-tetR-DT-(1)XbaI & PstI
6

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

Nameconcentration[µg/mL]260/280260/230
Pcon-RBS-GFP-DT (EcoRI&SpeI)7.11.880.07
Pcon-RBS-luxR-DT (EcoRI&XbaI)15.31.930.70
Plux (SpeI&PstI)4.51.790.31
RBS-GFP-DT (XbaI&PstI)6.02.160.03
RBS-tetR-DT (XbaI&PstI)7.61.910.38


Liquid Culture

No name

Samplemedium
8/21 RBS-Lysis2-9Plusgrow medium(+Amp)
8/21 RBS-Lysis2-12Plusgrow medium(+Amp)
8/21 RBS-Lysis2-14Plusgrow medium(+Amp)　

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

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