Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Gel Extraction)
(→Colony PCR)
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</div>
</div>
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===Colony PCR===
+
===Electrophoresis===
<div class="experiment">
<div class="experiment">
<span class="author">No name</span>
<span class="author">No name</span>
{| class="wikitable"
{| class="wikitable"
-
!|Sample||base pair
+
!Lane||Sample
|-
|-
-
|Pconst-spinach-DT-(1)||-
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|1||100bp ladder
|-
|-
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|Pconst-spinach-DT-(2)||-
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|2||Pconst-spinach-DT -(1)
|-
|-
-
|Pconst-spinach-DT-(3)||-
+
|3||Pconst-spinach-DT -(2)
|-
|-
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|Pconst-spinach-DT-(4)||-
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|4||Pconst-spinach-DT -(3)
|-
|-
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|Plac-spinach-DT-(1)||-
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|5||Pconst-spinach-DT -(4)
|-
|-
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|Plac-spinach-DT-(2)||-
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|6||Plac-spinach-DT -(1)
|-
|-
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|Plac-spinach-DT-(3)||-
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|7||Plac-spinach-DT -(2)
|-
|-
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|Plac-spinach-DT-(4)||-
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|8||Plac-spinach-DT -(3)
|-
|-
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|Pcon-pT181 antisense-(1)||-
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|9||Plac-spinach-DT -(4)
|-
|-
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|Pcon-pT181 antisense-(2)||-
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|10||Pcon-pT181 antisense -(1)
|-
|-
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|Pcon-pT181 antisense-(3)||-
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|11||Pcon-pT181 antisense -(2)
|-
|-
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|Pcon-pT181 antisense-(4)||-
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|12||Pcon-pT181 antisense -(3)
 +
|-
 +
|13||Pcon-pT181 antisense -(4)
|}
|}
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+
[[File:Igku_xxxxxx.xxx]]
{| class="wikitable"
{| class="wikitable"
-
!|Sample||base pair
+
!Lane||Sample
 +
|-
 +
|1||100bp ladder
|-
|-
-
|Ptet-pT181 antisense-(1)||-
+
|2||Ptet-pT181 antisense -(1)
|-
|-
-
|Ptet-pT181 antisense-(2)||-
+
|3||Ptet-pT181 antisense -(2)
|-
|-
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|Ptet-pT181 antisense-(3)||-
+
|4||Ptet-pT181 antisense -(3)
|-
|-
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|Ptet-pT181 antisense-(4)||-
+
|5||Ptet-pT181 antisense -(4)
|-
|-
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|Plac-pT181 attenuater-(1)||-
+
|6||Plac-pT181 attenuater -(1)
|-
|-
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|Plac-pT181 attenuater-(2)||-
+
|7||Plac-pT181 attenuater -(2)
|-
|-
-
|Plac-pT181 attenuater-(3)||-
+
|8||Plac-pT181 attenuater -(3)
|-
|-
-
|Plac-pT181 attenuater-(4)||-
+
|9||Plac-pT181 attenuater -(4)
|}
|}
 +
[[File:Igku_xxxxxx.xxx]]
===Miniprep===
===Miniprep===

Revision as of 15:43, 24 September 2013

Contents

Sep 6

Electrophoresis

Hirano

LaneSample
1100bp ladder
29/4 RBS-lysis3-DT -3
39/4 RBS-lysis3-DT -4
49/4 RBS-lysis3-DT -5
59/4 RBS-lysis1-DT -3
69/4 Ptrc-KaiC -1
79/4 pSB4K5 -1
8100bp ladder

File:Igku xxxxxx.xxx

Colony PCR

No name

Samplebase pair
Pconst-spinach-DT-(1~4)563
Plac-spinach-DT-(1~4)659
Pcon-pT181 antisense-(1~4)468
Ptet-pT181 antisense-(1~4)604
Pconst-pT181 attenuater-(1~4)568
Pconst+RBS-tetR-DT-(1~4)1121
Ptet+RBS-lacZα-DT-(1~4)1162
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s40s30cycles


Liquid Culture

No name

Samplemedium
9/4 RBS-Lysis3+DT3Plusgrow medium(+CP)
9/4 RBS-Lysis1+DT3Plusgrow medium(+CP)
9/4 pSB4K5Plusgrow medium(+Kan)

Electrophoresis

No name

LaneSample
1100bp ladder
2Pconst-spinach-DT -(1)
3Pconst-spinach-DT -(2)
4Pconst-spinach-DT -(3)
5Pconst-spinach-DT -(4)
6Plac-spinach-DT -(1)
7Plac-spinach-DT -(2)
8Plac-spinach-DT -(3)
9Plac-spinach-DT -(4)
10Pcon-pT181 antisense -(1)
11Pcon-pT181 antisense -(2)
12Pcon-pT181 antisense -(3)
13Pcon-pT181 antisense -(4)

File:Igku xxxxxx.xxx

LaneSample
1100bp ladder
2Ptet-pT181 antisense -(1)
3Ptet-pT181 antisense -(2)
4Ptet-pT181 antisense -(3)
5Ptet-pT181 antisense -(4)
6Plac-pT181 attenuater -(1)
7Plac-pT181 attenuater -(2)
8Plac-pT181 attenuater -(3)
9Plac-pT181 attenuater -(4)

File:Igku xxxxxx.xxx

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-luxIEcoRI & SpeI
3
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-luxI (EcoRI & SpeI)4.51.790.31

Liquid Culture

No name

Samplemedium
9/5　Pconst-spinach-DT-(1)Plusgrow medium(+Amp&CP)
9/5　Ptet-pT181 antisense-(1)Plusgrow medium(+CP)
9/5　Pconst+RBS-tetR-DT-(1)Plusgrow medium(+Amp&CP)
9/5　Ptet+RBS-lacZα-DT-(1)Plusgrow medium(+CP)
9/5　Pcon-pT181 attenuater-(1)Plusgrow medium(+CP)
9/4　pT181 antisense-(1)Plusgrow medium(+CP)
9/4　aptamer 12-1R-(1)Plusgrow medium(+CP)

Colony PCR

No name

Samplebase pair
9/5 Plac-spinach-DT-(5~12)563
9/5 Pcon-antisense-(5~12)468
9/5 Plac-attenuater-(5~12)664
9/5 pT181 antisense+Plac-(1)-
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s42s30cycles
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