Template:Kyoto/Notebook/Aug 22

From 2013.igem.org

(Difference between revisions)
(Created page with "==Aug 22== ===Liquid culture=== <div class="experiment"> <span class="author">Nakamoto</span> {| class="wikitable" !|Sample||Medium |- |8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (1)|...")
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===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">Tatsui, Kojima, Honda and Ashida</span>
 +
{| class="wikitable"
 +
!Fusion1 antisense (1) (300 &micro;g/mL)||EcoRI||SpeI||buffer H||MilliQ||total
 +
|-
 +
|3.3||1||1||3||21.7||30
 +
|-
 +
|0.7||0.2||0||1||8.1||10
 +
|-
 +
|0.7||0||0.2||1||8.1||10
 +
|-
 +
|0.7||0||0||1||8.3||10
 +
|}
 +
{| class="wikitable"
 +
!Fusion1 antisense (1) (300 &micro;g/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total
 +
|-
 +
|3.3||1||1||3||3||18.7||30
 +
|-
 +
|0.7||0.2||0||1||1||7.1||10
 +
|-
 +
|0.7||0||0.2||1||1||7.1||10
 +
|-
 +
|0.7||0||0||1||1||7.3||10
 +
|}
 +
{| class="wikitable"
 +
!Fusion6 antisense (1) (336 &micro;g/mL)||EcoRI||SpeI||buffer H||MilliQ||total
 +
|-
 +
|3||1||1||3||22||30
 +
|-
 +
|0.6||0.2||0||1||8.2||10
 +
|-
 +
|0.6||0||0.2||1||8.2||10
 +
|-
 +
|0.6||0||0||1||8.4||10
 +
|}
 +
{| class="wikitable"
 +
!Fusion6 antisense (1) (336 &micro;g/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total
 +
|-
 +
|3||1||1||3||3||19||30
 +
|-
 +
|0.6||0.2||0||1||1||7.2||10
 +
|-
 +
|0.6||0||0.2||1||1||7.2||10
 +
|-
 +
|0.6||0||0||1||1||7.4||10
 +
|}
 +
{| class="wikitable"
 +
!pT181 antisense (1) (178 &micro;g/mL)||EcoRI||SpeI||buffer H||MilliQ||total
 +
|-
 +
|5.6||1||1||3||19.4||30
 +
|-
 +
|1.1||0.2||0||1||7.7||10
 +
|-
 +
|1.1||0||0.2||1||7.7||10
 +
|-
 +
|1.1||0||0||1||7.9||10
 +
|}
 +
{| class="wikitable"
 +
!pT181 antisense (1) (178 &micro;g/mL)||XbaI||PstI||buffer M||BSA||MilliQ||total
 +
|-
 +
|5.6||1||1||3||3||16.4||30
 +
|-
 +
|1.1||0.2||0||1||1||6.7||10
 +
|-
 +
|1.1||0||0.2||1||1||6.7||10
 +
|-
 +
|1.1||0||0||1||1||6.9||10
 +
|}
 +
{| class="wikitable"
 +
!pT181 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total
 +
|-
 +
|3||1||1||3||22||30
 +
|-
 +
|0.6||0.2||0||1||8.2||10
 +
|-
 +
|0.6||0||0.2||1||8.2||10
 +
|-
 +
|0.6||0||0||1||8.4||10
 +
|}
 +
{| class="wikitable"
 +
!pT181 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total
 +
|-
 +
|3||1||1||3||3||19||30
 +
|-
 +
|0.6||0.2||0||1||1||7.2||10
 +
|-
 +
|0.6||0||0.2||1||1||7.2||10
 +
|-
 +
|0.6||0||0||1||1||7.4||10
 +
|}
 +
{| class="wikitable"
 +
!Fusion1 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total
 +
|-
 +
|3.3||1||1||3||21.7||30
 +
|-
 +
|0.7||0.2||0||1||8.1||10
 +
|-
 +
|0.7||0||0.2||1||8.1||10
 +
|-
 +
|0.7||0||0||1||8.3||10
 +
|}
 +
{| class="wikitable"
 +
!Fusion1 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total
 +
|-
 +
|3.3||1||1||3||3||18.7||30
 +
|-
 +
|0.7||0.2||0||1||1||7.1||10
 +
|-
 +
|0.7||0||0.2||1||1||7.1||10
 +
|-
 +
|0.7||0||0||1||1||7.3||10
 +
|}
 +
{| class="wikitable"
 +
!Fusion3m2 attenuator (1)||EcoRI||SpeI||buffer H||MilliQ||total
 +
|-
 +
|3||1||1||3||22||30
 +
|-
 +
|0.6||0.2||0||1||8.2||10
 +
|-
 +
|0.6||0||0.2||1||8.2||10
 +
|-
 +
|0.6||0||0||1||8.4||10
 +
|}
 +
{| class="wikitable"
 +
!Fusion3m2 attenuator (1)||XbaI||PstI||buffer M||BSA||MilliQ||total
 +
|-
 +
|3||1||1||3||3||19||30
 +
|-
 +
|0.6||0.2||0||1||1||7.2||10
 +
|-
 +
|0.6||0||0.2||1||1||7.2||10
 +
|-
 +
|0.6||0||0||1||1||7.4||10
 +
|}
 +
{| class="wikitable"
 +
!Spinach (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
 +
|-
 +
|2.3||0.5||0.5||3||0.3||23.4||30
 +
|-
 +
|0.5||0.1||0||1||0.1||8.3||10
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|-
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|0.5||0||0.1||1||0.1||8.3||10
 +
|-
 +
|0.5||0||0||1||0.1||8.4||10
 +
|}
 +
{| class="wikitable"
 +
!tetR aptamer 12_P (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
 +
|-
 +
|2.7||0.5||0.5||3||0.3||23||30
 +
|-
 +
|0.5||0.1||0||1||0.1||8.3||10
 +
|-
 +
|0.5||0||0.1||1||0.1||8.3||10
 +
|-
 +
|0.5||0||0||1||0.1||8.4||10
 +
|}
 +
{| class="wikitable"
 +
!tetR aptamer 12_1R (2)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
 +
|-
 +
|2.6||0.5||0.5||3||0.3||23.1||30
 +
|-
 +
|0.5||0.1||0||1||0.1||8.3||10
 +
|-
 +
|0.5||0||0.1||1||0.1||8.3||10
 +
|-
 +
|0.5||0||0||1||0.1||8.4||10
 +
|}
 +
{| class="wikitable"
 +
!tetR aptamer 12_1M (1)||EcoRI||SpeI||buffer B||BSA||MilliQ||total
 +
|-
 +
|2.7||0.5||0.5||3||0.3||23||30
 +
|-
 +
|0.5||0.1||0||1||0.1||8.3||10
 +
|-
 +
|0.5||0||0.1||1||0.1||8.3||10
 +
|-
 +
|0.5||0||0||1||0.1||8.4||10
 +
|}
 +
{| class="wikitable"
 +
!J23100||SpeI||PstI||buffer B||BSA||MilliQ||total
 +
|-
 +
|7||0.5||0.5||3||0.3||18.7||30
 +
|-
 +
|1.4||0.1||0||1||0.1||7.4||10
 +
|-
 +
|1.4||0||0.1||1||0.1||7.4||10
 +
|-
 +
|1.4||0||0||1||0.1||7.5||10
 +
|}
 +
{| class="wikitable"
 +
!Plac (2)||SpeI||PstI||buffer B||BSA||MilliQ||total
 +
|-
 +
|11.2||0.5||0.5||3||0.3||14.5||30
 +
|-
 +
|2.2||0.1||0||1||0.1||6.6||10
 +
|-
 +
|2.2||0||0.1||1||0.1||6.6||10
 +
|-
 +
|2.2||0||0||1||0.1||6.7||10
 +
|}
 +
 +
</div>
===Miniprep===
===Miniprep===
===Electrophoresis===
===Electrophoresis===

Revision as of 23:35, 16 September 2013

Contents

Aug 22

Liquid culture

Nakamoto

SampleMedium
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (1)8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (2)8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT control (1)8/21 Plusgrow medium (+Amp)
8/21 Pcon-RBS-GFP-DT control (2)8/21 Plusgrow medium (+Amp)
8/21 RBS-lysis1 (1)8/21 Plusgrow medium (+Amp)
8/21 RBS-lysis1 (2)8/21 Plusgrow medium (+Amp)
8/21 RBS control (1)8/21 Plusgrow medium (+Amp)
8/21 RBS control (2)8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm)-RBS-tetR-DT (1)8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm)-RBS-tetR-DT (2)8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm) control (1)8/21 Plusgrow medium (+Amp)
8/21 Ptet(pm) control (2)8/21 Plusgrow medium (+Amp)
8/21 Plux-RBS-GFP-DT (1)8/21 Plusgrow medium (+Amp)
8/21 pSB1C3(BBa_J04450) (1)8/21 Plusgrow medium (+Amp)
8/21 pSB1C3(BBa_J04450) (2)8/21 Plusgrow medium (+Amp)

incubate 37°C 10hour

Colony PCR

Kojima

Samplebase pair
8/21 RBS-lysis1(1)400
8/21 RBS-lysis1(2)400
8/21 RBS control(1)--
8/21 RBS control(2)--
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles
Samplebase pair
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (1)2143
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (2)2143
8/21 Pcon-RBS-luxR control(1)--
8/21 Pcon-RBS-luxR control(2)--
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s2min30cycles
Samplebase pair
8/21 Ptet-RBS-tetR-DT (1)1216
8/21 Ptet-RBS-tetR-DT (2)1216
8/21 Ptet control (1)--
8/21 Ptet control (2)--
8/21 Plux-RBS-GFP-DT (1)1227
8/21 pSB1C3(BBa_J04450) (1)1353
8/21 pSB1C3(BBa_J04450) (2)1353
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1pT181antisenseEcoRISpeI
2pT181antisenseEcoRI--
3pT181antisense--SpeI
4pT181antisense----
5pT181antisenseXbaIPstI
6pT181antisenseXbaI--
7pT181antisense--PstI
8pT181antisense----
9100bp ladder----
10Fusion1 attenuatorEcoRISpeI
11Fusion1 attenuatorEcoRI--
12Fusion1 attenuator--SpeI
13Fusion1 attenuator----
14Fusion1 attenuatorXbaIPstI
15Fusion1 attenuatorXbaI--
16fusion1 attenuator--PstI
17fusion1 attenuator----

File:Igku xxxxxx.xxx

LaneSampleEnzyme1Enzyme2
1Fusion3 3m2 attenuatorEcoRISpeI
2Fusion3 3m2 attenuatorEcoRI--
3Fusion3 3m2 attenuator--SpeI
4Fusion3 3m2 attenuator----
5Fusion3 3m2 attenuatorXbaIPstI
6Fusion3 3m2 attenuatorXbaI--
7Fusion3 3m2 attenuator--PstI
8Fusion3 3m2 attenuator----
9100bp ladder----
10aptamer 12-1MEcoRISpeI
11aptamer 12-1MEcoRI--
12aptamer 12-1M--SpeI
13aptamer 12-1M----
14aptamer 12-PEcoRISpeI
15aptamer 12-PEcoRI--
16aptamer 12-1P--SpeI
17aptamer 12-1P----

File:Igku xxxxxx.xxx

Restriction Enzyme Digestion

Tatsui, Kojima, Honda and Ashida

Fusion1 antisense (1) (300 µg/mL)EcoRISpeIbuffer HMilliQtotal
3.311321.730
0.70.2018.110
0.700.218.110
0.70018.310
Fusion1 antisense (1) (300 µg/mL)XbaIPstIbuffer MBSAMilliQtotal
3.3113318.730
0.70.20117.110
0.700.2117.110
0.700117.310
Fusion6 antisense (1) (336 µg/mL)EcoRISpeIbuffer HMilliQtotal
31132230
0.60.2018.210
0.600.218.210
0.60018.410
Fusion6 antisense (1) (336 µg/mL)XbaIPstIbuffer MBSAMilliQtotal
311331930
0.60.20117.210
0.600.2117.210
0.600117.410
pT181 antisense (1) (178 µg/mL)EcoRISpeIbuffer HMilliQtotal
5.611319.430
1.10.2017.710
1.100.217.710
1.10017.910
pT181 antisense (1) (178 µg/mL)XbaIPstIbuffer MBSAMilliQtotal
5.6113316.430
1.10.20116.710
1.100.2116.710
1.100116.910
pT181 attenuator (1)EcoRISpeIbuffer HMilliQtotal
31132230
0.60.2018.210
0.600.218.210
0.60018.410
pT181 attenuator (1)XbaIPstIbuffer MBSAMilliQtotal
311331930
0.60.20117.210
0.600.2117.210
0.600117.410
Fusion1 attenuator (1)EcoRISpeIbuffer HMilliQtotal
3.311321.730
0.70.2018.110
0.700.218.110
0.70018.310
Fusion1 attenuator (1)XbaIPstIbuffer MBSAMilliQtotal
3.3113318.730
0.70.20117.110
0.700.2117.110
0.700117.310
Fusion3m2 attenuator (1)EcoRISpeIbuffer HMilliQtotal
31132230
0.60.2018.210
0.600.218.210
0.60018.410
Fusion3m2 attenuator (1)XbaIPstIbuffer MBSAMilliQtotal
311331930
0.60.20117.210
0.600.2117.210
0.600117.410
Spinach (2)EcoRISpeIbuffer BBSAMilliQtotal
2.30.50.530.323.430
0.50.1010.18.310
0.500.110.18.310
0.50010.18.410
tetR aptamer 12_P (2)EcoRISpeIbuffer BBSAMilliQtotal
2.70.50.530.32330
0.50.1010.18.310
0.500.110.18.310
0.50010.18.410
tetR aptamer 12_1R (2)EcoRISpeIbuffer BBSAMilliQtotal
2.60.50.530.323.130
0.50.1010.18.310
0.500.110.18.310
0.50010.18.410
tetR aptamer 12_1M (1)EcoRISpeIbuffer BBSAMilliQtotal
2.70.50.530.32330
0.50.1010.18.310
0.500.110.18.310
0.50010.18.410
J23100SpeIPstIbuffer BBSAMilliQtotal
70.50.530.318.730
1.40.1010.17.410
1.400.110.17.410
1.40010.17.510
Plac (2)SpeIPstIbuffer BBSAMilliQtotal
11.20.50.530.314.530
2.20.1010.16.610
2.200.110.16.610
2.20010.16.710

Miniprep

Electrophoresis

Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Aug_22"