Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Colony PCR)
(→Gel Extraction)
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|2||rowspan="2"|8/20 RBS-lysis1-(1)||rowspan="2"|EcoRI & SpeI
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|2||rowspan="2"|RBS-lysis2-DT||rowspan="2"|XbaI & PstI
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|5||rowspan="2"|8/28 RBS-GFP-DT-(2)||rowspan="2"|XbaI & PstI
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|5||rowspan="2"|aptamer 12-1R-DT||rowspan="2"|XbaI & PstI
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{| class="wikitable"
{| class="wikitable"
!Lane||DNA||Enzyme
!Lane||DNA||Enzyme
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|1||100bp ladder||--
|1||100bp ladder||--
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|2||rowspan="2"|8/28 Plux||rowspan="2"|SpeI & PstI
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|2||rowspan="2"|Pλ-lux3||rowspan="2"|EcoRI & SpeI
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|4||--||--
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|5||rowspan="2"|8/20 RBS-tetR-DT-(1)||rowspan="2"|XbaI & PstI
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{| class="wikitable"
{| class="wikitable"
!Name||concentration[&micro;g/mL]||260/280||260/230
!Name||concentration[&micro;g/mL]||260/280||260/230
|-
|-
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|Pcon-RBS-GFP-DT (EcoRI&SpeI)||7.1||1.88||0.07
+
|RBS-lysis2-DT (XbaI&PstI)||7.1||1.88||0.07
-
|-
+
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|Pcon-RBS-luxR-DT (EcoRI&XbaI)||15.3||1.93||0.70
+
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|-
+
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|Plux (SpeI&PstI)||4.5||1.79||0.31
+
|-
|-
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|RBS-GFP-DT (XbaI&PstI)||6.0||2.16||0.03
+
|aptamer 12-1R-DT (XbaI&PstI)||15.3||1.93||0.70
|-
|-
-
|RBS-tetR-DT (XbaI&PstI)||7.6||1.91||0.38
+
|Pλ-lux3 (EcoRI&SpeI)||4.5||1.79||0.31
|}
|}
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===Liquid Culture===
===Liquid Culture===

Revision as of 10:18, 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

Honda and Kojima

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

Hirano

Samplebase pair
Pconst＋RBS-tetR-DT-(1)1121
Pconst＋RBS-tetR-DT-(2)1121
Pconst＋RBS-tetR-DT-(3)1121
Pconst＋RBS-tetR-DT-(4)1121
Ptet＋RBS-lacZα-DT-(1)1162
Ptet＋RBS-lacZα-DT-(2)1162
Ptet＋RBS-lacZα-DT-(3)1162
Ptet＋RBS-lacZα-DT-(4)1162

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder--
2RBS-lysis2-DTXbaI & PstI
3
4----
5aptamer 12-1R-DTXbaI & PstI
6
LaneDNAEnzyme
1100bp ladder--
2Pλ-lux3EcoRI & SpeI
3
4
5
6
Nameconcentration[µg/mL]260/280260/230
RBS-lysis2-DT (XbaI&PstI)7.11.880.07
aptamer 12-1R-DT (XbaI&PstI)15.31.930.70
Pλ-lux3 (EcoRI&SpeI)4.51.790.31

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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