Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

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Line 1: Line 1:
==Sep 6==
==Sep 6==
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/15 BBa_K112806(lysis2) -(1)||828
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||1min||30cycles
 +
|}
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|8/10 BBa_Q0400 -(1)||3995
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
 +
===Liquid Culture===
 +
<!-- ここから -->
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|(・ω・) ||
 +
|-
 +
|（・Ｘ・） ||　
 +
|}
 +
</div>
 +
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/15 BBa_K112806(lysis2) -(1)||828
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||1min||30cycles
 +
|}
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|8/10 BBa_Q0400 -(1)||3995
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
 +
===Miniprep===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{|class="wikitable"
 +
!DNA||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|(・▽・)||　||　||
 +
|}
 +
</div>
 +
<!-- ここまでをコピーしてね -->
 +
 +
===Restriction Enzyme Digestion===
 +
<div class="experiment">
 +
<span class="author">Murata, Murakami, Nakamoto, Ashida, Matsumoto and Yoshida</span>
 +
{| class="wikitable"
 +
! ||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
 +
|}
 +
*at 37&deg;C, for 1h
 +
{| 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
 +
|}
 +
*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
 +
{| class="wikitable"
 +
! ||8/17 K748002(lysis3) -(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 I13504(RBS-GFP-DT) -(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 I0462(RBS-luxR-DT) -(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 I732020(RBS-lacZ&alpha;-DT) -(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"
 +
! ||8/11 R6061 -(1)||SpeI||PstI||Buffer||MilliQ||total
 +
|-
 +
|2 cuts||8.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||17&micro;L||30&micro;L
 +
|-
 +
|1 cut||3.0&micro;L||0.2&micro;L||--||1.0&micro;L||5.8&micro;L||10&micro;L
 +
|-
 +
|1 cut||3.0&micro;L||--||0.2&micro;L||1.0&micro;L||5.8&micro;L||10&micro;L
 +
|-
 +
|NC||3.0&micro;L||--||--||1.0&micro;L||6.0&micro;L||10&micro;L
 +
|}
 +
*at 37&deg;C, for 1h
 +
</div>
 +
 +
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">Ashida</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100bp ladder||--
 +
|-
 +
|3||8/18 K117000(lysis2) -1||XbaI & PstI
 +
|-
 +
|5||8/18 K117000(lysis2) -1||XbaI & PstI
 +
|}
 +
[[File:igku_Aug19electrophoresis3]]<br>
 +
[[File:igku_Aug19electrophoresis4]]<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|8/18 lysis2-1(XbaI & PstI)||3.1||3.64||0.19
 +
|-
 +
|8/18 lysis2-1(XbaI & PstI)||3.4||1.89||0.13
 +
|}
 +
 +
</div>
 +
 +
 +
===Liquid Culture===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|8/15 BBa_K112806(lysis2) -(1)||Plusgrow medium(+CP)
 +
|-
 +
|8/15 BBa_Q04400(tetR) -(1)||Plusgrow medium(+Amp)
 +
|-
 +
|}
 +
</div>
 +
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/15 BBa_K112806(lysis2) -(1)||828
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||1min||30cycles
 +
|}
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|8/10 BBa_Q0400 -(1)||3995
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}
 +
 +
===Colony PCR===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!|Sample||base pair
 +
|-
 +
|8/15 BBa_K112806(lysis2) -(1)||828
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||1min||30cycles
 +
|}
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|8/10 BBa_Q0400 -(1)||3995
 +
|-
 +
|negative control||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||30s||30cycles
 +
|}

Revision as of 07:24, 24 September 2013

Contents

Sep 6

Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles
Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

Liquid Culture

No name

Samplemedium
(・ω・)
（・Ｘ・） 　


Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles
Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
(・▽・)　　

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 K748002(lysis3) -(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 I13504(RBS-GFP-DT) -(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 I0462(RBS-luxR-DT) -(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 I732020(RBS-lacZα-DT) -(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
8/11 R6061 -(1)SpeIPstIBufferMilliQtotal
2 cuts8.0µL1.0µL1.0µL3.0µL17µL30µL
1 cut3.0µL0.2µL--1.0µL5.8µL10µL
1 cut3.0µL--0.2µL1.0µL5.8µL10µL
NC3.0µL----1.0µL6.0µL10µL
  • at 37°C, for 1h

Gel Extraction

Ashida

LaneDNAEnzyme
1100bp ladder--
38/18 K117000(lysis2) -1XbaI & PstI
58/18 K117000(lysis2) -1XbaI & PstI

File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4

Nameconcentration[µg/mL]260/280260/230
8/18 lysis2-1(XbaI & PstI)3.13.640.19
8/18 lysis2-1(XbaI & PstI)3.41.890.13


Liquid Culture

Nakamoto

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


Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles
Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles
Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles
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