Template:Kyoto/Notebook/Sep 3

From 2013.igem.org

(Difference between revisions)
(→Electrophoresis)
(→Restriction Enzyme Digestion)
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<span class="author">Kozima</span>
<span class="author">Kozima</span>
{| class="wikitable"
{| class="wikitable"
-
! ||DNA||EcoR1||Spe1||Xba1||Pst1||10xBuffer||10xBuffer||100xBSA||MilliQ||total
+
! ||DNA||EcoR1||Spe1||Xba1||Pst1||10xBuffer B(EcoR1&Spe1)||10xBuffer D(Xba1&Pst1)||100xBSA||MilliQ||total
|-
|-
|pT181attenuater 14:00 418.4ng/&micro;L|| 4.8&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 0&micro;L|| 0.3&micro;L|| 19.9&micro;L|| 30&micro;L
|pT181attenuater 14:00 418.4ng/&micro;L|| 4.8&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 0&micro;L|| 0.3&micro;L|| 19.9&micro;L|| 30&micro;L
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|pT181attenuater 14:00 418.4ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
|pT181attenuater 14:00 418.4ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L
|}
|}
 +
</div>
 +
<div class="experiment">
 +
{| class="wikitable"
 +
! ||DNA||EcoR1||Spe1||Xba1||Pst1||10xBuffer B(EcoR1&Spe1)||10xBuffer D(Xba1&Pst1)||100xBSA||MilliQ||total
 +
|-
 +
|pT181antisense 14:00 362.2ng/&micro;L|| 5.5&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 0&micro;L|| 3&micro;L|| 0&micro;L|| 0.3&micro;L|| 19.2&micro;L|| 30&micro;L
 +
|-
 +
|pT181attenuater 14:00 418.4ng/&micro;L|| 0.3&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0&micro;L|| 0.1&micro;L|| 8.6&micro;L|| 10&micro;L
 +
|-
 +
|pT181attenuater 14:00 418.4ng/&micro;L|| 5.5&micro;L|| 0&micro;L|| 0&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 0&micro;L|| 3&micro;L|| 0.3&micro;L|| 19.2&micro;L|| 30&micro;L
 +
|-
 +
|pT181attenuater 14:00 418.4ng/&micro;L|| 0.3&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.6&micro;L|| 10&micro;L
 +
|}
 +
</div>
 +
 +
<div class="experiment">
 +
{| class="wikitable"
 +
! ||DNA||EcoR1||Spe1||10xBuffer B||100xBSA||MilliQ||total
 +
|-
 +
|apt 12_1R 14:00 507.5ng/&micro;L|| 3.9&micro;L|| 1.0&micro;L|| 1.0&micro;L|| 3&micro;L|| 0.3&micro;L|| 20.8&micro;L|| 30&micro;L
 +
|-
 +
|apt 12_1R 14:00 507.5ng/&micro;L|| 0.2&micro;L|| 0&micro;L|| 0&micro;L|| 1&micro;L|| 0.1&micro;L|| 8.7&micro;L|| 10&micro;L|}
</div>
</div>

Revision as of 10:20, 25 September 2013

Contents

Sep 3

Electrophoresis

Kozima

LaneSample
1100bp ladder
2pT181 attenuater(2)-DT2(738)
3Ptet-RBS-lacZα-DT-2(756)
4Ptet-RBS-lacZα-DT-3
5Ptet-RBS-lacZα-DT-4
6Ptet-RBS-lacZα-DT-5
7Ptet-RBS-lacZα-DT-6
8RBS-lysis2-DT-2(985)
9RBS-lysis2-DT-3
10100bp ladder

Igku Sep3 Electrophoresis(N1).jpg

Miniprep

Tatsui

DNAconcentration[µg/mL]260/280260/230
9/2 14:00 pT181 attenuator418.41.821.80
9/2 14:00 pT181 antisense362.21.921.96
9/2 14:00 Spinach438.31.861.99
9/2 14:00 tetR-apt 12_1R567.51.901.99
9/2 21:00 pT181 attenuater320.91.902.15
9/2 21:00 Spinach457.01.882.24

Liquid Culture

Kozima

Samplemedium
Spinach-DT(9/2 master plate)plusgrow(CP)4ml

Restriction Enzyme Digestion

Kozima

DNAEcoR1Spe1Xba1Pst110xBuffer B(EcoR1&Spe1)10xBuffer D(Xba1&Pst1)100xBSAMilliQtotal
pT181attenuater 14:00 418.4ng/µL 4.8µL 1.0µL 1.0µL 0µL 0µL 3µL 0µL 0.3µL 19.9µL 30µL
pT181attenuater 14:00 418.4ng/µL 0.2µL 0µL 0µL 0µL 0µL 1µL 0µL 0.1µL 8.7µL 10µL
pT181attenuater 14:00 418.4ng/µL 4.8µL 0µL 0µL 1.0µL 1.0µL 0µL 3µL 0.3µL 19.9µL 30µL
pT181attenuater 14:00 418.4ng/µL 0.2µL 0µL 0µL 0µL 0µL 0µL 1µL 0.1µL 8.7µL 10µL
DNAEcoR1Spe1Xba1Pst110xBuffer B(EcoR1&Spe1)10xBuffer D(Xba1&Pst1)100xBSAMilliQtotal
pT181antisense 14:00 362.2ng/µL 5.5µL 1.0µL 1.0µL 0µL 0µL 3µL 0µL 0.3µL 19.2µL 30µL
pT181attenuater 14:00 418.4ng/µL 0.3µL 0µL 0µL 0µL 0µL 1µL 0µL 0.1µL 8.6µL 10µL
pT181attenuater 14:00 418.4ng/µL 5.5µL 0µL 0µL 1.0µL 1.0µL 0µL 3µL 0.3µL 19.2µL 30µL
pT181attenuater 14:00 418.4ng/µL 0.3µL 0µL 0µL 0µL 0µL 0µL 1µL 0.1µL 8.6µL 10µL
DNAEcoR1Spe110xBuffer B100xBSAMilliQtotal
apt 12_1R 14:00 507.5ng/µL 3.9µL 1.0µL 1.0µL 3µL 0.3µL 20.8µL 30µL
apt 12_1R 14:00 507.5ng/µL 0.2µL 0µL 0µL 1µL 0.1µL 8.7µL}

PCR

Mutatation PCR

Colony PCR

Restriction Enzyme Digestion

Gel Extraction

Liquid Culture
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