Template:Kyoto/Notebook/Aug 18

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==Aug 18==
==Aug 18==
-
==Gel Extraction==
+
===Gel Extraction===
 +
<div class="experiment">
 +
<span class="author">Yoshida and Kojima</span>
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|1||100kb ladder||--
 +
|-
 +
|2||8/11 J23100 -&#9313;||--
 +
|-
 +
|3||8/11 J23100 -&#9313;||--
 +
|-
 +
|4||--||--
 +
|-
 +
|5||8/17 K117000(lysis1) -1||--
 +
|-
 +
|6||8/17 K117000(lysis1) -1||--
 +
|-
 +
|7||--||--
 +
|-
 +
|8||8/17 K748002(lysis3) -1||--
 +
|-
 +
|9||8/17 K748002(lysis3) -1||--
 +
|-
 +
|10||--||--
 +
|-
 +
|11||8/17 I13504(RBS-GFP-DT) -1||--
 +
|-
 +
|12||8/17 I13504(RBS-GFP-DT) -1||--
 +
|}
 +
[[File:Igku Aug18 Gel Extraction before.jpg]]<br>
 +
[[File:Igku Aug18 Gel Extraction1-2.jpg]]<br>
 +
 
 +
{| class="wikitable"
 +
!Lane||DNA||Enzyme
 +
|-
 +
|13||1kb ladder||--
 +
|-
 +
|14||8/17 I0462(RBS-luxR-DT) -1||--
 +
|-
 +
|15||8/17 I0462(RBS-luxR-DT) -1||--
 +
|-
 +
|16||--||--
 +
|-
 +
|17||8/17 I732020(RBS-lacZ&alpha;-DT) -1||--
 +
|-
 +
|18||8/17 I732020(RBS-lacZ&alpha;-DT) -1||--
 +
|}
 +
[[File:Igku Aug18 Gel Extraction2 before.jpg]]<br>
 +
[[File:Igku Aug18 Gel Extraction2-2.jpg]]<br>
 +
<br>
 +
{| class="wikitable"
 +
!Name||concentration[&micro;g/mL]||260/280||260/230
 +
|-
 +
|8/14 J231000||1.7||1.61||0.04
 +
|-
 +
|8/17 K748002(lysis3) -1||5.4||2.39||0.45
 +
|-
 +
|8/17 I13504(RBS-GFP-DT) -1||3.4||3.33||0.19
 +
|-
 +
|8/17 I0462(RBS-luxR-DT) -1||4.4||2.07||0.37
 +
|-
 +
|8/17 I732020(RBS-lacZ&alpha;-DT) -1||2.9||2.17||0.29
 +
|}
 +
</div>
 +
 
 +
===Ligation===
 +
<div class="experiment">
 +
<span class="author">Ashida</span>
 +
{| class="wikitable"
 +
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
 +
|-
 +
|experiment||J23100||9.0||RBS-lacZα-DT||8.6||3.5
 +
|-
 +
|experiment||J23100||9.0||RBS-GFP-DT||7.3||3.5
 +
|-
 +
|experiment||RBS ||1||lysis3||4.6||2.8
 +
|-
 +
|experiment||Plux ||1||GFP||7.3||3.5
 +
|-
 +
|experiment||J23100 ||9.0||--||--||3.5
 +
|-
 +
|experiment||RBS||1.0||--||--||0.5
 +
|-
 +
|experiment||Plux||1.0||--||--||0.5
 +
|}
 +
incubate 16&deg;C 1hour
 +
</div>
 +
 
 +
===Restriction Enzyme Digestion===
 +
<!-- こっから -->
 +
<div class="experiment">
 +
<span class="author">Murata and Okazaki</span>
 +
{| class="wikitable"
 +
! ||8/17K112806(lysis2)-1||XbaI||PstI||Buffer||BSA||MilliQ||total
 +
|-
 +
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L|| 3.0&micro;L||30 &micro;L
 +
|-
 +
|1 cuts||1.0&mic6/8ro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L|| 6.8&micro;L||10 &micro;L
 +
|-
 +
|1 cuts||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
 +
|}
 +
</div>
 +
 
 +
===Electrophoresis===
 +
 +
<!-- こっから -->
 +
<div class="experiment">
 +
<span class="author">Okazaki and Honda</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||--||--
 +
|-
 +
|2||lysis1 ||XbaI||PstI
 +
|-
 +
|3||lysis1||XbaI||--
 +
|-
 +
|4||lysis1||--||PstI
 +
|-
 +
|5||lysis1||--||--
 +
|-
 +
|6||lysis2||XbaI||PstI
 +
|-
 +
|7||lysis2||XbaI||--
 +
|-
 +
|8||lysis2||--||PstI
 +
|-
 +
|9||lysis2||--||--
 +
|-
 +
|10||100bp ladder||--||--
 +
|}
 +
[[File:Igku_Aug18electrophoresis.jpg]]<br>
 +
</div>
 +
 
 +
===Colony PCR===
 +
<!-- ここから -->
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||base pair
 +
|-
 +
|8/8 LBKana||--
 +
|-
 +
|8/16 Pbad araC||--
 +
|-
 +
|NC||--
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 +
|-
 +
|5min||30s||30s||1min||30
 +
|}
 +
</div>
 +
 
 +
===Liquid Culture===
 +
<!-- ここから -->
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|7/8 LB Kana ||Plusgrow(+Kana)
 +
|-
 +
|8/16 Pbad araC||Plusgrow(+Kana)
 +
|}
 +
</div>
 +
 
 +
===LB Medium Plate===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!volume||200ml
 +
|-
 +
|Bacto Trypton||2g
 +
|-
 +
|Bacto yeasy extract||1g
 +
|-
 +
|NaCl||1g
 +
|-
 +
|Agar Powder||2g
 +
|}
 +
</div>
 +
 
 +
===Transformation===
 +
<div class="experiment">
 +
<span class="author">No name </span>
 +
{| class="wikitable"
 +
!Name||Sample||Competent Cells||Total||Plate
 +
|-
 +
|8/18 J23100+RBS-LacZ&alpha;-DT||2&micro;L||20&micro;L||22&micro;L||Amp
 +
|-
 +
|8/18 J23100+RBS-GFP-DT||2&micro;L||20&micro;L||22&micro;L||Amp
 +
|-
 +
|8/18 J23100+RBS-luxR-DT ||2&micro;L||20&micro;L||22&micro;L||Amp
 +
|-
 +
|RBS+lysis3||2&micro;L||20&micro;L||22&micro;L||Amp
 +
|-
 +
|Plux+GFP||2&micro;L||20&micro;L||22&micro;L||CP
 +
|-
 +
|J23100||2&micro;L||20&micro;L||22&micro;L||Amp
 +
|-
 +
|RBS||2&micro;L||20&micro;L||22&micro;L||Amp
 +
|-
 +
|Plux||2&micro;L||20&micro;L||22&micro;L||CP
 +
|}
 +
</div>
 +
<!-- ここまでをコピーしてね -->
 +
 
 +
===Electrophoresis===
 +
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||-- ||--
 +
|-
 +
|2||8/16Pbad araC-1||--||--
 +
|-
 +
|3||8/16Pbad araC-2||--||--
 +
|-
 +
|4||NC||--||--
 +
|}
 +
[[File:Igku Aug18electrophoresis2.jpg]]<br>
 +
</div>

Latest revision as of 03:43, 28 September 2013

Contents

Aug 18

Gel Extraction

Yoshida and Kojima

LaneDNAEnzyme
1100kb ladder--
28/11 J23100 -②--
38/11 J23100 -②--
4----
58/17 K117000(lysis1) -1--
68/17 K117000(lysis1) -1--
7----
88/17 K748002(lysis3) -1--
98/17 K748002(lysis3) -1--
10----
118/17 I13504(RBS-GFP-DT) -1--
128/17 I13504(RBS-GFP-DT) -1--

Igku Aug18 Gel Extraction before.jpg
Igku Aug18 Gel Extraction1-2.jpg

LaneDNAEnzyme
131kb ladder--
148/17 I0462(RBS-luxR-DT) -1--
158/17 I0462(RBS-luxR-DT) -1--
16----
178/17 I732020(RBS-lacZα-DT) -1--
188/17 I732020(RBS-lacZα-DT) -1--

Igku Aug18 Gel Extraction2 before.jpg
Igku Aug18 Gel Extraction2-2.jpg

Nameconcentration[µg/mL]260/280260/230
8/14 J2310001.71.610.04
8/17 K748002(lysis3) -15.42.390.45
8/17 I13504(RBS-GFP-DT) -13.43.330.19
8/17 I0462(RBS-luxR-DT) -14.42.070.37
8/17 I732020(RBS-lacZα-DT) -12.92.170.29

Ligation

Ashida

stateVectorInserterLigation High ver.2
experimentJ231009.0RBS-lacZα-DT8.63.5
experimentJ231009.0RBS-GFP-DT7.33.5
experimentRBS 1lysis34.62.8
experimentPlux 1GFP7.33.5
experimentJ23100 9.0----3.5
experimentRBS1.0----0.5
experimentPlux1.0----0.5

incubate 16°C 1hour

Restriction Enzyme Digestion

Murata and Okazaki

8/17K112806(lysis2)-1XbaIPstIBufferBSAMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL 3.0µL30 µL
1 cuts1.0&mic6/8ro;L0.2µL--1.0µL1.0µL 6.8µL10 µL
1 cuts1.0µL--0.2µL1.0µL1.0µL 6.8µL10 µL

Electrophoresis

Okazaki and Honda

LaneSampleEnzyme1Enzyme2
1100bp ladder----
2lysis1 XbaIPstI
3lysis1XbaI--
4lysis1--PstI
5lysis1----
6lysis2XbaIPstI
7lysis2XbaI--
8lysis2--PstI
9lysis2----
10100bp ladder----

Igku Aug18electrophoresis.jpg

Colony PCR

No name

Samplebase pair
8/8 LBKana--
8/16 Pbad araC--
NC--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30

Liquid Culture

No name

Samplemedium
7/8 LB Kana Plusgrow(+Kana)
8/16 Pbad araCPlusgrow(+Kana)

LB Medium Plate

No name

volume200ml
Bacto Trypton2g
Bacto yeasy extract1g
NaCl1g
Agar Powder2g

Transformation

No name

NameSampleCompetent CellsTotalPlate
8/18 J23100+RBS-LacZα-DT2µL20µL22µLAmp
8/18 J23100+RBS-GFP-DT2µL20µL22µLAmp
8/18 J23100+RBS-luxR-DT 2µL20µL22µLAmp
RBS+lysis32µL20µL22µLAmp
Plux+GFP2µL20µL22µLCP
J231002µL20µL22µLAmp
RBS2µL20µL22µLAmp
Plux2µL20µL22µLCP

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder-- --
28/16Pbad araC-1----
38/16Pbad araC-2----
4NC----

Igku Aug18electrophoresis2.jpg

Retrieved from "http://2013.igem.org/Template:Kyoto/Notebook/Aug_18"