Template:Kyoto/Notebook/Aug 18

From 2013.igem.org

(Difference between revisions)
(→Aug 18)
(→Liquid Culture)
 
(18 intermediate revisions not shown)
Line 8: Line 8:
|1||100kb ladder||--
|1||100kb ladder||--
|-
|-
-
|2||8/11 J23100-②||--
+
|2||8/11 J23100 -②||--
|-
|-
-
|3||8/11 J23100-②||--
+
|3||8/11 J23100 -②||--
|-
|-
|4||--||--
|4||--||--
|-
|-
-
|5||8/17 K117000()lysis1)-1||--
+
|5||8/17 K117000(lysis1) -1||--
|-
|-
-
|6||8/17 K117000()lysis1)-1||--
+
|6||8/17 K117000(lysis1) -1||--
|-
|-
|7||--||--
|7||--||--
|-
|-
-
|8||8/17 K748002(lysis3)-1||--
+
|8||8/17 K748002(lysis3) -1||--
|-
|-
-
|9||8/17 K748002(lysis3)-1||--
+
|9||8/17 K748002(lysis3) -1||--
|-
|-
|10||--||--
|10||--||--
|-
|-
-
|11||8/17 I13504(RBS-GFP-DT)-1||--
+
|11||8/17 I13504(RBS-GFP-DT) -1||--
|-
|-
-
|12||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||--
|13||1kb ladder||--
|-
|-
-
|14||8/17 I0462(RBS-luxR-DT)-1||--
+
|14||8/17 I0462(RBS-luxR-DT) -1||--
|-
|-
-
|15||8/17 I0462(RBS-luxR-DT)-1||--
+
|15||8/17 I0462(RBS-luxR-DT) -1||--
|-
|-
|16||--||--
|16||--||--
|-
|-
-
|17||8/17 I732020(RBS-lacZ&alpha;-DT)-1||--
+
|17||8/17 I732020(RBS-lacZ&alpha;-DT) -1||--
|-
|-
-
|18||8/17 I732020(RBS-lacZ&alpha;-DT)-1||--
+
|18||8/17 I732020(RBS-lacZ&alpha;-DT) -1||--
|}
|}
-
[fig before]
+
[[File:Igku Aug18 Gel Extraction2 before.jpg]]<br>
-
[fig after]
+
[[File:Igku Aug18 Gel Extraction2-2.jpg]]<br>
 +
<br>
{| class="wikitable"
{| class="wikitable"
!Name||concentration[&micro;g/mL]||260/280||260/230
!Name||concentration[&micro;g/mL]||260/280||260/230
Line 49: Line 56:
|8/14 J231000||1.7||1.61||0.04
|8/14 J231000||1.7||1.61||0.04
|-
|-
-
|8/17 K748002(lysis3)-1||5.4||2.39||0.45
+
|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 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 I0462(RBS-luxR-DT) -1||4.4||2.07||0.37
|-
|-
-
|8/17 I732020(RBS-lacZ-DT)-1||2.9||2.17||0.29
+
|8/17 I732020(RBS-lacZ&alpha;-DT) -1||2.9||2.17||0.29
|}
|}
</div>
</div>
Line 65: Line 72:
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
|-
|-
-
|experiment||J23100||9.0||RBS-lacZα-DT||8.6||
+
|experiment||J23100||9.0||RBS-lacZα-DT||8.6||3.5
|-
|-
-
|experiment||J23100||9.0||RBS-GFP-DT||7.3||
+
|experiment||J23100||9.0||RBS-GFP-DT||7.3||3.5
|-
|-
-
|experiment||RBS ||1||lysis3||4.6||
+
|experiment||RBS ||1||lysis3||4.6||2.8
|-
|-
-
|experiment||Plux ||1||GFP||7.3||
+
|experiment||Plux ||1||GFP||7.3||3.5
|-
|-
-
|experiment||J23100 ||9.0||--||--||
+
|experiment||J23100 ||9.0||--||--||3.5
|-
|-
-
|experiment||RBS||1.0||--||--||
+
|experiment||RBS||1.0||--||--||0.5
|-
|-
-
|experiment||Plux||1.0||--||--||
+
|experiment||Plux||1.0||--||--||0.5
|}
|}
incubate 16&deg;C 1hour  
incubate 16&deg;C 1hour  
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|1||100bp ladder||--||--
|1||100bp ladder||--||--
|-  
|-  
-
|2||lysis1 ||Xba1||Pst1
+
|2||lysis1 ||XbaI||PstI
|-
|-
-
|3||lysis1||Xba1||--
+
|3||lysis1||XbaI||--
|-
|-
-
|4||lysis1||--||Pst1
+
|4||lysis1||--||PstI
|-
|-
|5||lysis1||--||--
|5||lysis1||--||--
|-
|-
-
|6||lysis2||Xba1||Pst1
+
|6||lysis2||XbaI||PstI
|-
|-
-
|7||lysis2||Xba1||--
+
|7||lysis2||XbaI||--
|-
|-
-
|8||lysis2||--||Pst1
+
|8||lysis2||--||PstI
|-
|-
|9||lysis2||--||--
|9||lysis2||--||--
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|10||100bp ladder||--||--
|10||100bp ladder||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:Igku_Aug18electrophoresis.jpg]]<br>
</div>
</div>
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|8/16 Pbad araC||--
|8/16 Pbad araC||--
|-
|-
-
|negative control||--
+
|NC||--
|}
|}
{| class="wikitable"
{| class="wikitable"
Line 147: Line 154:
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
|-
|-
-
|5min||30s||30s||1min||
+
|5min||30s||30s||1min||30
|}
|}
</div>
</div>
Line 160: Line 167:
|7/8 LB Kana ||Plusgrow(+Kana)
|7/8 LB Kana ||Plusgrow(+Kana)
|-
|-
-
|8/16 Pbad araC||Plusgrow(+Kana)　
+
|8/16 Pbad araC||Plusgrow(+Kana)
|}
|}
</div>
</div>
-
===Medium Plate===
+
===LB Medium Plate===
<div class="experiment">
<div class="experiment">
<span class="author">No name</span>
<span class="author">No name</span>
Line 174: Line 181:
|Bacto yeasy extract||1g
|Bacto yeasy extract||1g
|-
|-
-
|NaCl\\1g
+
|NaCl||1g
|-
|-
|Agar Powder||2g
|Agar Powder||2g
Line 207: Line 214:
===Electrophoresis===
===Electrophoresis===
   
   
-
<!-- こっから -->
 
<div class="experiment">
<div class="experiment">
<span class="author">No name</span>
<span class="author">No name</span>
Line 219: Line 225:
|3||8/16Pbad araC-2||--||--
|3||8/16Pbad araC-2||--||--
|-
|-
-
|4||negative control||--||--
+
|4||NC||--||--
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:Igku Aug18electrophoresis2.jpg]]<br>
</div>
</div>
-
<!-- ここまでをコピーしてね -->
 

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

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