Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Sep 6)
(→Sep 6)
Line 17: Line 17:
|Pconst-PT181 attenuater-(1~4)||568
|Pconst-PT181 attenuater-(1~4)||568
|-
|-
-
|Pconst＋RBS-tetR-DT-(1~4)||1121
+
|Pconst+RBS-tetR-DT-(1~4)||1121
|-
|-
-
|Ptet＋RBS-lacZα-DT-(1~4)||1162
+
|Ptet+RBS-lacZα-DT-(1~4)||1162
|}
|}
{| class="wikitable"
{| class="wikitable"
Line 37: Line 37:
!Sample||medium
!Sample||medium
|-
|-
-
|9/4 RBS-Lysis3＋DT3||Plusgrow medium(+Cp)
+
|9/4 RBS-Lysis3+DT3||Plusgrow medium(+Cp)
|-
|-
-
|9/4 RBS-Lysis1＋DT3||Plusgrow medium(+Cp)
+
|9/4 RBS-Lysis1+DT3||Plusgrow medium(+Cp)
|-
|-
|9/4 PSB4K5||Plusgrow medium(+K)　
|9/4 PSB4K5||Plusgrow medium(+K)　
Line 120: Line 120:
|PSB4K5||212.9||1.80||1.90
|PSB4K5||212.9||1.80||1.90
|-
|-
-
|RBS-lacZα-DT||185.7||1.78||1.79
+
|RBS-lacZα-DT||185.7||1.78||1.79
|-
|-
|RBS-lysis2-DT-(1)||84.8||1.85||2.24
|RBS-lysis2-DT-(1)||84.8||1.85||2.24
Line 162: Line 162:
!|Sample||base pair
!|Sample||base pair
|-
|-
-
|Pconst＋RBS-tetR-DT-(1)||1121
+
|Pconst+RBS-tetR-DT-(1)||1121
|-
|-
-
|Pconst＋RBS-tetR-DT-(2)||1121
+
|Pconst+RBS-tetR-DT-(2)||1121
|-
|-
-
|Pconst＋RBS-tetR-DT-(3)||1121
+
|Pconst+RBS-tetR-DT-(3)||1121
|-
|-
-
|Pconst＋RBS-tetR-DT-(4)||1121
+
|Pconst+RBS-tetR-DT-(4)||1121
|-
|-
-
|Ptet＋RBS-lacZα-DT-(1)||1162
+
|Ptet+RBS-lacZα-DT-(1)||1162
|-
|-
-
|Ptet＋RBS-lacZα-DT-(2)||1162
+
|Ptet+RBS-lacZα-DT-(2)||1162
|-
|-
-
|Ptet＋RBS-lacZα-DT-(3)||1162
+
|Ptet+RBS-lacZα-DT-(3)||1162
|-
|-
-
|Ptet＋RBS-lacZα-DT-(4)||1162
+
|Ptet+RBS-lacZα-DT-(4)||1162
|}
|}
</div>
</div>
Line 234: Line 234:
|9/5　Ptet-PT181 antisense-(1)||Plusgrow medium(+CP)
|9/5　Ptet-PT181 antisense-(1)||Plusgrow medium(+CP)
|-
|-
-
|9/5　Pconst＋RBS-tetR-DT-(1)||Plusgrow medium(+Amp&CP)
+
|9/5　Pconst+RBS-tetR-DT-(1)||Plusgrow medium(+Amp&CP)
|-
|-
-
|9/5　Ptet＋RBS-lacZα-DT-(1)||Plusgrow medium(+CP)
+
|9/5　Ptet+RBS-lacZ&alpha;-DT-(1)||Plusgrow medium(+CP)
|-
|-
|9/5　Pcon-PT181 attenuater-(1)||Plusgrow medium(+CP)
|9/5　Pcon-PT181 attenuater-(1)||Plusgrow medium(+CP)
Line 258: Line 258:
|9/5 Plac-attenuater-(5~12)||664
|9/5 Plac-attenuater-(5~12)||664
|-
|-
-
|9/5 PT181 antisense＋Plac-(1)||-
+
|9/5 PT181 antisense+Plac-(1)||-
|}
|}
{| class="wikitable"
{| class="wikitable"

Revision as of 15:13, 24 September 2013

Contents

Sep 6

Electrophoresis

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(+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
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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