Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Liquid Culture)
(→Gel Extraction)
Line 218: Line 218:
|4||--||--
|4||--||--
|-
|-
-
|5||rowspan="2"|aptamer 12-1R-DT||rowspan="2"|XbaI & PstI
+
|5||rowspan="2"|aptamer 12_1R-DT||rowspan="2"|XbaI & PstI
|-
|-
|6
|6
Line 227: Line 227:
|1||100bp ladder||--
|1||100bp ladder||--
|-
|-
-
|2||rowspan="2"|Pλ-lux3||rowspan="2"|EcoRI & SpeI
+
|2||rowspan="2"|Pλ-luxI||rowspan="2"|EcoRI & SpeI
|-
|-
|3
|3
-
|-
 
-
|4
 
-
|-
 
-
|5
 
-
|-
 
-
|6
 
|}
|}
{| class="wikitable"
{| class="wikitable"
!Name||concentration[µg/mL]||260/280||260/230
!Name||concentration[µg/mL]||260/280||260/230
|-
|-
-
|RBS-lysis2-DT (XbaI&PstI)||7.1||1.88||0.07
+
|RBS-lysis2-DT (XbaI & PstI)||7.1||1.88||0.07
|-
|-
-
|aptamer 12-1R-DT (XbaI&PstI)||15.3||1.93||0.70
+
|aptamer 12_1R-DT (XbaI & PstI)||15.3||1.93||0.70
|-
|-
-
|Pλ-lux3 (EcoRI&SpeI)||4.5||1.79||0.31
+
|Pλ-luxI (EcoRI & SpeI)||4.5||1.79||0.31
|}
|}
</div>
</div>

Revision as of 15:35, 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)

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-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
Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Sep_6"