Template:Kyoto/Notebook/Aug 17

From 2013.igem.org

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</div>
</div>
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===Restriction Enzyme Digestion===
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<div class="experiment">
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<span class="author">Murata,Murakami,Nakamoto,Ashida,Matsumoto and Yoshida</span>
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{| class="wikitable"
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! ||8/11 J23100 -(2)||SPeI||PstI||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||4.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||21&micro;L||30&micro;L
 +
|-
 +
|1 cut||1.3&micro;L||0.2&micro;L||--||1.0&micro;L||7.5&micro;L||10&micro;L
 +
|-
 +
|1 cut||1.3&micro;L||--||0.2&micro;L||1.0&micro;L||7.5&micro;L||10&micro;L
 +
|-
 +
|NC||1.3&micro;L||--||--||1.0&micro;L||7.7&micro;L||10&micro;L
 +
|}
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*at 37&deg;C, for 1h
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{| class="wikitable"
 +
! ||8/15 B0034(RBS) -(2)||SPeI||PstI||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||6.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||19&micro;L||30&micro;L
 +
|-
 +
|1 cut||2.0&micro;L||0.2&micro;L||--||1.0&micro;L||6.8&micro;L||10&micro;L
 +
|-
 +
|1 cut||2.0&micro;L||--||0.2&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 +
|-
 +
|NC||2.0&micro;L||--||--||1.0&micro;L||7.0&micro;L||10&micro;L
 +
|}
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*at 37&deg;C, for 1h
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{| class="wikitable"
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! ||8/17 K117000(lysis1) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L||20&micro;L||30&micro;L
 +
|-
 +
|1 cut||1.0&micro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 +
|-
 +
|1 cut||1.0&micro;L||--||0.2&micro;L||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 +
|-
 +
|NC||1.0&micro;L||--||--||1.0&micro;L||1.0&micro;L||7.0&micro;L||10&micro;L
 +
|}
 +
*at 37&deg;C, for 1h
 +
{| class="wikitable"
 +
! ||8/17 K117000(lysis1) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L||20&micro;L||30&micro;L
 +
|-
 +
|1 cut||1.0&micro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 +
|-
 +
|1 cut||1.0&micro;L||--||0.2&micro;L||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 +
|-
 +
|NC||1.0&micro;L||--||--||1.0&micro;L||1.0&micro;L||7.0&micro;L||10&micro;L
 +
|}
 +
*at 37&deg;C, for 1h
 +
</div>
 +
===Master Plate===
===Master Plate===
<div class="experiment">
<div class="experiment">

Revision as of 15:49, 23 September 2013

Contents

Aug 17

Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles

File:Image

Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

File:Image

Liquid Culture

Nakamoto

Samplemedium
8/15 BBa_K112806(lysis2) -(1)Plusgrow medium(+CP)
8/15 BBa_Q04400(tetR) -(1)Plusgrow medium(+Amp)

Restriction Enzyme Digestion

Murata,Murakami,Nakamoto,Ashida,Matsumoto and Yoshida

8/11 J23100 -(2)SPeIPstIBufferMilliQtotal
2 cuts4.0µL1.0µL1.0µL3.0µL21µL30µL
1 cut1.3µL0.2µL--1.0µL7.5µL10µL
1 cut1.3µL--0.2µL1.0µL7.5µL10µL
NC1.3µL----1.0µL7.7µL10µL
  • at 37°C, for 1h
8/15 B0034(RBS) -(2)SPeIPstIBufferMilliQtotal
2 cuts6.0µL1.0µL1.0µL3.0µL19µL30µL
1 cut2.0µL0.2µL--1.0µL6.8µL10µL
1 cut2.0µL--0.2µL1.0µL6.8µL10µL
NC2.0µL----1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 K117000(lysis1) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 K117000(lysis1) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h

Master Plate

Nakamoto

NumberUse LB plate (+CP)
1BBa_K112806(lysis2) -(1)
NumberUse LB plate (+Kana)
1BBa_Q04400(tetR) -(1)
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