Template:Kyoto/Notebook/Sep 15

From 2013.igem.org

(Difference between revisions)
(→Restriction Enzyme Digestion)
(→Liquid Culture)
 
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Line 108: Line 108:
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
|-
|-
-
|1,14 || || ||  
+
|1,14 ||100bp ladder|| ||  
|-
|-
|2 ||Pcon-RBS-tetR-DT ||EcoRI ||SpeI  
|2 ||Pcon-RBS-tetR-DT ||EcoRI ||SpeI  
Line 122: Line 122:
|7 ||Pcon-apt12-1R-DT || ||  
|7 ||Pcon-apt12-1R-DT || ||  
|-
|-
-
|8 ||Pcon-spin-DT ||EcoRI ||SpeI
+
|8 ||Pcon-spinach-DT ||EcoRI ||SpeI
|-
|-
-
|9 ||Pcon-spin-DT || ||  
+
|9 ||Pcon-spinach-DT || ||  
|-
|-
-
|10 ||Pcon-pT181 antisense-spin-DT ||XbaI ||PstI
+
|10 ||Pcon-pT181 antisense-spinach-DT ||XbaI ||PstI
|-
|-
-
|11 ||Pcon-pT181 antisense-spin-DT || ||  
+
|11 ||Pcon-pT181 antisense-spinach-DT || ||  
|-
|-
|12 ||RBS-lacZα-DT ||XbaI ||PstI
|12 ||RBS-lacZα-DT ||XbaI ||PstI
Line 140: Line 140:
|17 ||1kbp ladder || ||  
|17 ||1kbp ladder || ||  
|}
|}
-
[[File:igku_xxxxxx.xxx]]<br>
+
[[File:igku_9151.jpg]]<br>
</div>
</div>
<!-- ここまでをコピーしてね -->
<!-- ここまでをコピーしてね -->
 +
===Gel Extraction===
===Gel Extraction===
<div class="experiment">
<div class="experiment">
Line 156: Line 157:
|6~8 ||Pcon-apt12_1R-DT ||
|6~8 ||Pcon-apt12_1R-DT ||
|-
|-
-
|10~12 ||Pcon-spin-DT ||
+
|10~12 ||Pcon-spinach-DT ||
|}
|}
 +
[[File:igku_9152.jpg]]<br>
 +
[[File:igku_9153.jpg]]<br>
2
2
{| class="wikitable"
{| class="wikitable"
Line 164: Line 167:
|1 ||100bp ladder ||
|1 ||100bp ladder ||
|-
|-
-
|2~4 ||Pcon-pT181 anticodone-spin-DT ||XbaI & PstI
+
|2~4 ||Pcon-pT181 anticodone-spinach-DT ||XbaI & PstI
|-
|-
|6~8 ||RBS-lacZ&alpha;-DT ||XbaI & PstI
|6~8 ||RBS-lacZ&alpha;-DT ||XbaI & PstI
Line 171: Line 174:
|}
|}
-
[[File:igku_xxbeforexx.xxx]]<br>
+
[[File:igku_9154.jpg]]<br>
-
[[File:igku_xxafterxx.xxx]]<br>
+
[[File:igku_9155.jpg]]<br>
{| class="wikitable"
{| class="wikitable"
!Name||concentration[&micro;g/mL]||260/280||260/230
!Name||concentration[&micro;g/mL]||260/280||260/230
Line 182: Line 185:
|9/15 RBS-lacZ&alpha;-DT (XbaI & PstI) ||1.0 ||0.85 ||0.44
|9/15 RBS-lacZ&alpha;-DT (XbaI & PstI) ||1.0 ||0.85 ||0.44
|-
|-
-
|Pcon-pT181 anticodone-spin-DT (XbaI & PstI) ||1.3 ||0.95 ||0.28
+
|Pcon-pT181 anticodone-spinach-DT (XbaI & PstI) ||1.3 ||0.95 ||0.28
|-
|-
-
|Pcon-spin-DT ||0.9 ||1.37 ||0.05
+
|Pcon-spinach-DT ||0.9 ||1.37 ||0.05
|-
|-
|Pcon-apt12_1R-DT ||1.7 ||1.42 ||0.29
|Pcon-apt12_1R-DT ||1.7 ||1.42 ||0.29
Line 191: Line 194:
|}
|}
</div>
</div>
 +
===Ligation===
===Ligation===
   
   
Line 200: Line 204:
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-RBS-tetR-DT EcoRI & SpeI||1.9 &micro;L||xx &micro;L
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-RBS-tetR-DT EcoRI & SpeI||1.9 &micro;L||xx &micro;L
|-
|-
-
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-spin-DT EcoRI & SpeI||4.7 &micro;L||xx &micro;L
+
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-spinach-DT EcoRI & SpeI||4.7 &micro;L||xx &micro;L
|-
|-
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-apt12_1R-DT EcoRI & SpeI||2.0 &micro;L||xx &micro;L
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-apt12_1R-DT EcoRI & SpeI||2.0 &micro;L||xx &micro;L
Line 206: Line 210:
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-pT181 attenuator-DT EcoRI & SpeI||3.0 &micro;L||xx &micro;L
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/15 Pcon-pT181 attenuator-DT EcoRI & SpeI||3.0 &micro;L||xx &micro;L
|-
|-
-
|experiment||9/13 pSB1C3 XbaI & PstI||1.9 &micro;L||9/15 Pcon-pT181 anticodone-spin-DT XbaI & PstI||4.4 &micro;L||xx &micro;L
+
|experiment||9/13 pSB1C3 XbaI & PstI||1.9 &micro;L||9/15 Pcon-pT181 anticodone-spinach-DT XbaI & PstI||4.4 &micro;L||xx &micro;L
|-
|-
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/3 pT181 attenuator EcoRI & SpeI||3.8 &micro;L||xx &micro;L
|experiment||9/13 pSB1C3 EcoRI & SpeI||4.7 &micro;L||9/3 pT181 attenuator EcoRI & SpeI||3.8 &micro;L||xx &micro;L
Line 212: Line 216:
|experiment||9/6 Pcon SpeI & PstI||1.4 &micro;L||9/15 RBS-lacZ&alpha;-DT XbaI & PstI||2.0 &micro;L||xx &micro;L
|experiment||9/6 Pcon SpeI & PstI||1.4 &micro;L||9/15 RBS-lacZ&alpha;-DT XbaI & PstI||2.0 &micro;L||xx &micro;L
|-
|-
-
|experiment||9/14 pSB 4k5 EcoRI & SpeI|| &micro;L||9/15 Pcon-spin-DT EcoRI & SpeI||9.4 &micro;L||xx &micro;L
+
|experiment||9/14 pSB 4k5 EcoRI & SpeI|| &micro;L||9/15 Pcon-spinach-DT EcoRI & SpeI||9.4 &micro;L||xx &micro;L
|-
|-
|experiment||9/15 Peet-RBS-lacZ&alpha;-DT EcoRI & XbaI||2.6 &micro;L||Pcon-RBS-tetR-DT EcoRI & SpeI||1.5 &micro;L||xx &micro;L
|experiment||9/15 Peet-RBS-lacZ&alpha;-DT EcoRI & XbaI||2.6 &micro;L||Pcon-RBS-tetR-DT EcoRI & SpeI||1.5 &micro;L||xx &micro;L
Line 218: Line 222:
|}
|}
</div>
</div>
 +
===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
<div class="experiment">
<div class="experiment">
Line 259: Line 264:
! ||DNA||SpeI||PstI||Buffer||BSA||MilliQ||total
! ||DNA||SpeI||PstI||Buffer||BSA||MilliQ||total
|-
|-
-
|9/10 spinarch-DT SpeI & PstI||9.9 &micro;L||1 &micro;L||1 &micro;L||3 &micro;L||3 &micro;L||12.1 &micro;L||30 &micro;L
+
|9/10 spinach-DT SpeI & PstI||9.9 &micro;L||1 &micro;L||1 &micro;L||3 &micro;L||3 &micro;L||12.1 &micro;L||30 &micro;L
|-
|-
|NC||0.5 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.5 &micro;L||10 &micro;L
|NC||0.5 &micro;L||0 &micro;L||0 &micro;L||1 &micro;L||1 &micro;L||7.5 &micro;L||10 &micro;L
Line 265: Line 270:
</div>
</div>
 +
===Liquid Culture===
===Liquid Culture===
-
<!-- ここから -->
 
<div class="experiment">
<div class="experiment">
<span class="author">No name</span>
<span class="author">No name</span>
Line 276: Line 281:
|DT ||
|DT ||
|-
|-
-
|Spinarch-DT || 　
+
|Spinach-DT || 　
|}
|}
</div>
</div>
-
<!-- ここまでをコピーしてね -->
 

Latest revision as of 00:39, 27 September 2013

Contents

Sep 15

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
Pλ-luxI225　1.59　0.83

Liquid Culture

No name

Samplemedium
9/13 Pλ-luxI-1
9/7 pSB4K5-1
9/11 Plac-3

Ligation

No name

stateVectorInserterLigation High ver.2
experiment9/13 Pcon(SpeI & PstI)3.6 µL9/12 RBS-lacZα-DT(XbaI & PstI)4.7 µL8 µL

Restriction Enzyme Digestion

No Name

DNAEcorISpeIBufferBSAMilliQtotal
9/7Pcon-RBS-tetR-DT (EcoRI & SpeI)15 µL1 µL1 µL3 µL3 µL6 µL30 µL
NC0.7 µL0 µL0 µL1 µL1 µL7.3 µL10 µL


DNAEcoRISpeIBufferBSAMilliQtotal
9/12 Pcon-pT181 attenuator-DT (EcoRI & SpeI)6.8 µL1 µL1 µL3 µL3 µL15.2 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAEcoRISpeIBufferBSAMilliQtotal
9/12 Pcon-apt12_1R-DT(EcoRI & SpeI)6.6 µL1 µL1 µL3 µL3 µL15.4 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAEcoRISpeIBufferBSAMilliQtotal
9/8 Pcon-spinach-DT (EcoRI & SpeI)7.6 µL1 µL1 µL3 µL3 µL15.4 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
9/11 Pcon-pT181 anticodone-spinach-DT(XbaI & PstI)7.2 µL1 µL1 µL3 µL3 µL14.8 µL30 µL
NC0.4 µL0 µL0 µL1 µL1 µL7.6 µL10 µL


DNAXbaIPstIBufferBSAMilliQtotal
8/17 RBS-lacZα-DT (XbaI & PstI)5.3 µL1 µL1 µL3 µL3 µL16.7 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL


DNAEcoRIXbaIBufferBSAMilliQtotal
9/7 Ptet-RBS-LacZα-DT (EcoRI & XbaI)2.9 µL1 µL1 µL3 µL3 µL18.5 µL30 µL
NC1.5 µL0 µL0 µL1 µL1 µL7 µL10 µL


Electrophoresis

No Name

LaneSampleEnzyme1Enzyme2
1,14 100bp ladder
2 Pcon-RBS-tetR-DT EcoRI SpeI
3 Pcon-RBS-tetR-DT
4 Pcon-pT181 attenuator-DT EcoRI SpeI
5 Pcon-pT181 attenuator-DT
6 Pcon-apt12-1R-DT EcoRI SpeI
7 Pcon-apt12-1R-DT
8 Pcon-spinach-DT EcoRI SpeI
9 Pcon-spinach-DT
10 Pcon-pT181 antisense-spinach-DT XbaI PstI
11 Pcon-pT181 antisense-spinach-DT
12 RBS-lacZα-DT XbaI PstI
13 RBS-lacZα-DT
15 Ptet-RBS-lacZα-DT EcoRI XbaI
16 Ptet-RBS-lacZα-DT
17 1kbp ladder

Igku 9151.jpg

Gel Extraction

No Name 1

LaneDNAEnzyme
1 100bp ladder
2~4 Pcon-pT181 attenuator-DT EcoRI & SpeI
6~8 Pcon-apt12_1R-DT
10~12 Pcon-spinach-DT

Igku 9152.jpg
Igku 9153.jpg
2

LaneDNAEnzyme
1 100bp ladder
2~4 Pcon-pT181 anticodone-spinach-DT XbaI & PstI
6~8 RBS-lacZα-DT XbaI & PstI
10~12 Pcon-RBS-tetR-DT EcoRI & SpeI

Igku 9154.jpg
Igku 9155.jpg

Nameconcentration[µg/mL]260/280260/230
9/15 Ptet-RBS-lacZα-DT (EcoRI & SpeI) 1.9 1.08 0.77
9/15 Pcon-RBS-tetR-DT (EcoRI & SpeI) 5.9 1.30 0.17
9/15 RBS-lacZα-DT (XbaI & PstI) 1.0 0.85 0.44
Pcon-pT181 anticodone-spinach-DT (XbaI & PstI) 1.3 0.95 0.28
Pcon-spinach-DT 0.9 1.37 0.05
Pcon-apt12_1R-DT 1.7 1.42 0.29
Pcon-pT181 attenuator-DT (EcoRI & SpeI) 2.1 1.53 0.46

Ligation

No Name

stateVectorInserterLigation High ver.2
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-RBS-tetR-DT EcoRI & SpeI1.9 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-spinach-DT EcoRI & SpeI4.7 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-apt12_1R-DT EcoRI & SpeI2.0 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/15 Pcon-pT181 attenuator-DT EcoRI & SpeI3.0 µLxx µL
experiment9/13 pSB1C3 XbaI & PstI1.9 µL9/15 Pcon-pT181 anticodone-spinach-DT XbaI & PstI4.4 µLxx µL
experiment9/13 pSB1C3 EcoRI & SpeI4.7 µL9/3 pT181 attenuator EcoRI & SpeI3.8 µLxx µL
experiment9/6 Pcon SpeI & PstI1.4 µL9/15 RBS-lacZα-DT XbaI & PstI2.0 µLxx µL
experiment9/14 pSB 4k5 EcoRI & SpeI µL9/15 Pcon-spinach-DT EcoRI & SpeI9.4 µLxx µL
experiment9/15 Peet-RBS-lacZα-DT EcoRI & XbaI2.6 µLPcon-RBS-tetR-DT EcoRI & SpeI1.5 µLxx µL

Restriction Enzyme Digestion

No Name

DNAXbaIPstIBufferBSAMilliQtotal
9/13 pT181 attenuator XbaI & PstI6.3 µL1 µL1 µL3 µL3 µL15.7 µL30 µL
NC0.3 µL0 µL0 µL1 µL1 µL7.7 µL10 µL
DNAEcoRISpeIBufferBSAMilliQtotal
9/10 pSB1C3 EcoRI & SpeI8.1 µL1 µL1 µL3 µL3 µL13.9 µL30 µL
NC0.4 µL0 µL0 µL1 µL1 µL7.6 µL10 µL
DNAXbaIPstIBufferBSAMilliQtotal
9/14 pSB4k5 XbaI & PstI9.5 µL1 µL1 µL3 µL3 µL12.5 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL
DNASpeIPstIBufferBSAMilliQtotal
9/11 DT SpeI & PstI10.4 µL1 µL1 µL3 µL3 µL11.6 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL
DNASpeIPstIBufferBSAMilliQtotal
9/6 apt12_1R-DT SpeI & PstI7.9 µL1 µL1 µL3 µL3 µL14.1 µL30 µL
NC0.4 µL0 µL0 µL1 µL1 µL7.6 µL10 µL
DNASpeIPstIBufferBSAMilliQtotal
9/10 spinach-DT SpeI & PstI9.9 µL1 µL1 µL3 µL3 µL12.1 µL30 µL
NC0.5 µL0 µL0 µL1 µL1 µL7.5 µL10 µL

Liquid Culture

No name

Samplemedium
pSB1C3
DT
Spinach-DT 　
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