Template:Kyoto/Notebook/Aug 28

From 2013.igem.org

(Difference between revisions)
(→Colony PCR: edtor:kojima)
(→Colony PCR: by kojima)
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!|Sample||base pair
!|Sample||base pair
|-
|-
-
|8/21 RBS-lysis2-(3)||772
+
|8/27 pT181 attenuator-DT-1||738
|-
|-
-
|8/21 RBS-lysis2-(4)||772
+
|8/27 pT181 attenuator-DT-2||738
|-
|-
-
|8/21 RBS-lysis2-(5)||772
+
|8/27 pT181 attenuator-DT-3||738
|-
|-
-
|8/21 RBS-lysis2-(6)||772
+
|8/27 TetR-aptamer 12_1R-DT-1||521
 +
|-
 +
|8/27 TetR-aptamer 12_1R-DT-2||521
 +
|-
 +
|8/27 pT181 antisense-DT-1||542
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94°C||94°C||55°C||68°C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
[[File:igku_Aug28electrophoresis3?.png]]
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/27 Pλ-luxI-1||???
 +
|-
 +
|8/27 Pλ-luxI||???
|}
|}
{| class="wikitable"
{| class="wikitable"
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|94°C||94°C||55°C||68°C||--
|94°C||94°C||55°C||68°C||--
|-
|-
-
|5min||30s||30s||48s||30cycles
+
|5min||30s||30s||30min||3cycles
|}
|}
[[File:igku_Aug19electrophoresis1.png]]
[[File:igku_Aug19electrophoresis1.png]]

Revision as of 07:55, 21 September 2013

Contents

Aug 28

Electrophoresis

No name

LaneSample
1100bp
2SasA
3RpaA
4RpaB
5PkaiBC
6PCR NC
7100bp

File:Igku xxxxxx.xxx

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
8/27 Plux173.81.951.45

Ligation

No name

stateVectorInserter
experiment8/71 DT-13.08.0
  • Samples were evaporeted used evaporator into about 3 µL.
sampleMilliQLigation Hightotal
343.510.5
  • incubate one hour at 16 °C

Gel Extraction

No name

LaneDNA
1100bpladder
3SasA(8/27 Genome PCR production)
5RpaA(8/27 Genome PCR production)
7RpaB(8/27 Genome PCR production)
9PkaiBC(8/27 Genome PCR production)

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

Colony PCR

No name

Samplebase pair
8/21 RBS-lysis2-(3)772
8/21 RBS-lysis2-(4)772
8/21 RBS-lysis2-(5)772
8/21 RBS-lysis2-(6)772
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s48s30cycles

File:Igku Aug19electrophoresis1.png

Restriction Enzyme Digestion

No name

8/22 RBS-lysis1-(1)EcoRISpeIBufferBBSAMilliQtotal
2 cuts9µL0.5µL0.5µL3µL0.3µL16.7µL30µL
NC1µL0µL0µL1µL0.1µL7.9µL10µL
8/17 RBS-GFP-DT-(2)XbaIPstIBufferDBSAMilliQtotal
2 cuts4µL0.5µL0.5µL3µL0.3µL21.7µL30µL
NC0.5µL0µL0µL1µL0.1µL8.4µL10µL
8/20 Pcon-RBS-GFP-DT-(1)EcoRISpeIBufferBBSAMilliQtotal
2 cuts3µL0.5µL0.5µL3µL0.3µL22.7µL30µL
NC0.3µL0µL0µL1µL0.1µL8.6µL10µL
8/20 Pcon-RBS-luxR-DT-(1)EcoRIXbaIBufferDBSAMilliQtotal
2 cuts2µL0.5µL0.5µL3µL0.3µL23.7µL30µL
NC0.3µL0µL0µL1µL0.1µL8.6µL10µL

No name

8/28 PluxSpeIPstIBufferDBSAMilliQtotal
2 cuts6µL0.5µL0.5µL3µL0.3µL19.7µL30µL
NC1µL0µL0µL1µL0.1µL7.9µL10µL
8/20 RBS-tetR-DT-(1)XbaIPstIBufferDBSAMilliQtotal
2 cuts5µL0.5µL0.5µL3µL0.3µL20.7µL30µL
NC0.5µL0µL0µL1µL0.1µL8.4µL10µL
  • incubated 37°C 1hour

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
18/22 RBS-lysis1-(1)EcoRISpeI
28/22 RBS-lysis1-(1)----
38/17 RBS-GFP-DT-(2)XbaIPstI
48/17 RBS-GFP-DT-(2)----
58/20 Pcon-RBS-GFP-DT-(1)EcoRIXbaI
68/20 Pcon-RBS-GFP-DT-(1)----
7100bp ladder----
88/20 Pcon-RBS-luxR-DT-(1)EcoRIXbaI
98/20 Pcon-RBS-luxR-DT-(1)----
108/28 PluxSpeIPstI
118/28 Plux----
128/20 RBS-tetR-DTXbaIPstI
138/20 RBS-tetR-DT----

File:Igku Aug28electrophoresis1

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

Colony PCR

No name

Samplebase pair
8/27 pT181 attenuator-DT-1738
8/27 pT181 attenuator-DT-2738
8/27 pT181 attenuator-DT-3738
8/27 TetR-aptamer 12_1R-DT-1521
8/27 TetR-aptamer 12_1R-DT-2521
8/27 pT181 antisense-DT-1542
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug28electrophoresis3?.png

Samplebase pair
8/27 Pλ-luxI-1???
8/27 Pλ-luxI???
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30min3cycles

File:Igku Aug19electrophoresis1.png

Liquid Culture

Ligation

===Liquid Culture===
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