Template:Kyoto/Notebook/Aug 29

From 2013.igem.org

(Difference between revisions)
(→Liquid Culture)
(→Restriction Enzyme Digestion)
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===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
! ||8/22 psB1C3-(2)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total
 +
|-
 +
|2 cuts||8&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||0&micro;L||0.3&micro;L||17.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|-
 +
|2 cuts||8&micro;L||0&micro;L||0&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||3&micro;L||0.3&micro;L||17.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 tRMA-spinach(1)||XbaI||SpeI||BufferB||BSA||MilliQ||total
 +
|-
 +
|2 cuts||4&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||21.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.4&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 tetR aptamer12-1R(1)||EcoRI||SpeI||BufferB||BSA||MilliQ||total
 +
|-
 +
|2 cuts||3&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||22.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.5&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 PT181 attenuator-(2)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total
 +
|-
 +
|2 cuts||4&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||0&micro;L||0.3&micro;L||21.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0&micro;L||0.1&micro;L||8.4&micro;L||10&micro;L
 +
|-
 +
|2 cuts||4&micro;L||0&micro;L||0&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||3&micro;L||0.3&micro;L||21.7&micro;L||30&micro;L
 +
|-
 +
|NC||0.5&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.4&micro;L||10&micro;L
 +
|}
 +
{| class="wikitable"
 +
! ||8/21 PT181 antisense-(2)||EcoRI||SpeI||XbaI||PstI||BufferB||BufferD||BSA||MilliQ||total
 +
|-
 +
|2 cuts||10&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||0&micro;L||3&micro;L||0&micro;L||0.3&micro;L||15.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|-
 +
|2 cuts||10&micro;L||0&micro;L||0&micro;L||0.5&micro;L||0.5&micro;L||0&micro;L||3&micro;L||0.3&micro;L||15.7&micro;L||30&micro;L
 +
|-
 +
|NC||1&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||7.9&micro;L||10&micro;L
 +
|}
 +
*incubated 37&deg;C 1hour
 +
</div>
 +
===Transformation===
===Transformation===
===M9 Medium Plate===
===M9 Medium Plate===

Revision as of 10:53, 21 September 2013

Contents

Aug 29

Colony PCR

No name

Samplebase pair
8/27 Pcon-pT181 antisense-(1)542
8/26 pT181 attenuator-DT-(1)738
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Igku Aug19electrophoresis1.png

Electrophoresis

No name

LaneSample
1100bp ladder
28/21 RBS-lysis2-9(Colony PCR prodution)
38/21 RBS-lysis2-8(Colony PCR prodution)
48/21 RBS-lysis2-7(Colony PCR prodution)
58/21 RBS-lysis2-10(Colony PCR prodution)
68/21 RBS-lysis2-11(Colony PCR prodution)
78/21 RBS-lysis2-12(Colony PCR prodution)
88/21 RBS-lysis2-13(Colony PCR prodution)
98/21 RBS-lysis2-14(Colony PCR prodution)

File:Igku xxxxxx.xxx

miniprep

No Name

DNAconcentration [µg/mL]260/280260/230
Plac153.72.032.10
Pλ-luxⅠ-(1)76.52.072.06
Pλ-luxⅠ-(2)86.42.082.10
DT166.21.652.12
J23100190.11.682.34

Liquid Culture

No name

Samplemedium
8/21 RBS-Lysis2-9Plusgrow medium(+Amp)
8/21 RBS-Lysis2-12Plusgrow medium(+Amp)
8/21 RBS-Lysis2-14Plusgrow medium(+Amp)　

Restriction Enzyme Digestion

No name

8/22 psB1C3-(2)EcoRISpeIXbaIPstIBufferBBufferDBSAMilliQtotal
2 cuts8µL0.5µL0.5µL0µL0µL3µL0µL0.3µL17.7µL30µL
NC1µL0µL0µL0µL0µL1µL0µL0.1µL7.9µL10µL
2 cuts8µL0µL0µL0.5µL0.5µL0µL3µL0.3µL17.7µL30µL
NC1µL0µL0µL0µL0µL0µL1µL0.1µL7.9µL10µL
8/21 tRMA-spinach(1)XbaISpeIBufferBBSAMilliQtotal
2 cuts4µL0.5µL0.5µL3µL0.3µL21.7µL30µL
NC0.5µL0µL0µL1µL0.1µL8.4µL10µL
8/21 tetR aptamer12-1R(1)EcoRISpeIBufferBBSAMilliQtotal
2 cuts3µL0.5µL0.5µL3µL0.3µL22.7µL30µL
NC0.4µL0µL0µL1µL0.1µL8.5µL10µL
8/21 PT181 attenuator-(2)EcoRISpeIXbaIPstIBufferBBufferDBSAMilliQtotal
2 cuts4µL0.5µL0.5µL0µL0µL3µL0µL0.3µL21.7µL30µL
NC0.5µL0µL0µL0µL0µL1µL0µL0.1µL8.4µL10µL
2 cuts4µL0µL0µL0.5µL0.5µL0µL3µL0.3µL21.7µL30µL
NC0.5µL0µL0µL0µL0µL0µL1µL0.1µL8.4µL10µL
8/21 PT181 antisense-(2)EcoRISpeIXbaIPstIBufferBBufferDBSAMilliQtotal
2 cuts10µL0.5µL0.5µL0µL0µL3µL0µL0.3µL15.7µL30µL
NC1µL0µL0µL0µL0µL1µL0µL0.1µL7.9µL10µL
2 cuts10µL0µL0µL0.5µL0.5µL0µL3µL0.3µL15.7µL30µL
NC1µL0µL0µL0µL0µL0µL1µL0.1µL7.9µL10µL
  • incubated 37°C 1hour

Transformation

M9 Medium Plate

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