Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Colony PCR)
(→Miniprep)
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!DNA||concentration [µg/mL]||260/280||260/230
!DNA||concentration [µg/mL]||260/280||260/230
|-
|-
-
|Plac||153.7||2.03||2.10
+
|PT181 antisense-(1)||238.7||1.68||1.44
|-
|-
-
|Pλ-luxⅠ-(1)||76.5||2.07||2.06
+
|PT181 antisense-(2)||248.8||1.83||1.77
|-
|-
-
|Pλ-luxⅠ-(2)||86.4||2.08||2.10
+
|PT181 antisense-(3)||223.2||1.78||1.61
|-
|-
-
|DT||166.2||1.65||2.12
+
|PT181 antisense-(4)||220.8||1.89||1.92
|-
|-
-
|J23100||190.1||1.68||2.34
+
|tetR aptamer-(1)||252.5||1.89||1.90
 +
|-
 +
|tetR aptamer-(2)||211.2||1.90||1.99
 +
|-
 +
|DT181 attenuater||102.2||1.98||2.17
 +
|-
 +
|121R aptamer-DT||251.7||1.92||2.01
 +
|-
 +
|PSB4K5||212.9||1.80||1.90
 +
|-
 +
|RBS-lacZα-DT||185.7||1.78||1.79
 +
|-
 +
|RBS-lysis2-DT-(1)||84.8||1.85||2.24
|}
|}
</div>
</div>

Revision as of 09:49, 24 September 2013

Contents

Sep 6

Liquid Culture

No name

Samplemedium
9/4 RBS-Lysis3＋DT3Plusgrow medium(+Cp)
9/4 RBS-Lysis1＋DT3Plusgrow medium(+Cp)
9/4 PSB4K5Plusgrow medium(+K)　

Colony PCR

No name

Samplebase pair
Pconst-spinach-DT-(1)-
Pconst-spinach-DT-(2)-
Pconst-spinach-DT-(3)-
Pconst-spinach-DT-(4)-
Plac-spinach-DT-(1)-
Plac-spinach-DT-(2)-
Plac-spinach-DT-(3)-
Plac-spinach-DT-(4)-
Pcon-PT181 antisense-(1)-
Pcon-PT181 antisense-(2)-
Pcon-PT181 antisense-(3)-
Pcon-PT181 antisense-(4)-
Samplebase pair
Ptet-PT181 antisense-(1)-
Ptet-PT181 antisense-(2)-
Ptet-PT181 antisense-(3)-
Ptet-PT181 antisense-(4)-
Plac-PT181 attenuater-(1)-
Plac-PT181 attenuater-(2)-
Plac-PT181 attenuater-(3)-
Plac-PT181 attenuater-(4)-

Miniprep

No Name

DNAconcentration [µg/mL]260/280260/230
PT181 antisense-(1)238.71.681.44
PT181 antisense-(2)248.81.831.77
PT181 antisense-(3)223.21.781.61
PT181 antisense-(4)220.81.891.92
tetR aptamer-(1)252.51.891.90
tetR aptamer-(2)211.21.901.99
DT181 attenuater102.21.982.17
121R aptamer-DT251.71.922.01
PSB4K5212.91.801.90
RBS-lacZα-DT185.71.781.79
RBS-lysis2-DT-(1)84.81.852.24

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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