Template:Kyoto/Notebook/Aug 31

From 2013.igem.org

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|8||--||--
|8||--||--
|-
|-
-
|9||rowspan=2"|pSB1C3(1)||rowspan="2"|XbaI&PstI
+
|9||rowspan="2"|pSB1C3(1)||rowspan="2"|XbaI & PstI
 +
|-
 +
|10
|}
|}
-
[[File:igku_xxbeforexx.xxx]]<br>
+
[[File:Igku Aug30 Gel Extraction(N1).jpg]]<br>
-
[[File:igku_xxafterxx.xxx]]<br>
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[[File:Igku Aug30 Gel Extraction(N2-1).jpg]]<br>
{| class="wikitable"
{| class="wikitable"
!Lane||DNA||Enzyme
!Lane||DNA||Enzyme
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|11
|11
|}
|}
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[[File:igku_xxbeforexx.xxx]]<br>
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[[File:Igku Aug31 Gel Extraction(N2-2).jpg]]<br>
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[[File:igku_xxafterxx.xxx]]<br>
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[[File:Igku Aug31 Gel Extraction(N2-3).jpg]]<br>
{| class="wikitable"
{| class="wikitable"
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|6
|6
|}
|}
 +
[[File:Igku Aug31 Gel Extraction(N3-1).jpg]]<br>
 +
[[File:Igku Aug31 Gel Extraction(N3-2).jpg]]<br>
 +
{| class="wikitable"
{| class="wikitable"
!Name||concentration[&micro;g/mL]||260/280||260/230
!Name||concentration[&micro;g/mL]||260/280||260/230
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|tetR aptamer 12_1R(EcoRI&SpeI)||--||--||--
|tetR aptamer 12_1R(EcoRI&SpeI)||--||--||--
|}
|}
 +
===Ligation===  
===Ligation===  
<div class="experiment">
<div class="experiment">
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!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
|-
|-
-
|experiment||8/30 pSB1C3(EcoRI&SpeI)||6.6||8/30 tRNA-Spinach(EcoRI&SpeI)||12.5||3.5
+
|experiment||8/30 pSB1C3 (EcoRI & SpeI)||6.6||8/30 tRNA-Spinach (EcoRI & SpeI)||12.5||3.5
|-
|-
-
|experiment||8/30 pSB1C3(EcoRI&SpeI)||6.6||8/30 pT181 antisense(EcoRI&SpeI)||12.5||3.5
+
|experiment||8/30 pSB1C3 (EcoRI & SpeI)||6.6||8/30 pT181 antisense (EcoRI & SpeI)||12.5||3.5
 +
|-
 +
|experiment||8/30 pSB1C3 (XbaI & Pst)||12.5||8/30 pT181 attenuator (XbaI & PstI)||25||3.5
 +
|-
 +
|experiment||8/30 pSB1C3 (XbaI & Pst)||12.5||8/30 pT181 antisense (XbaI & PstI)||25||3.5
 +
|-
 +
|experiment||8/20 DT (EcoRI & XbaI)||6.6||8/30 pT181 attenuator(2) (EcoRI & SpeI)||12.5||3.5
 +
|-
 +
|experiment||8/20 DT (EcoRI & XbaI)||6.6||8/30 pT181 antisense (EcoRI & SpeI)||12.5||3.5
 +
|-
 +
|experiment||8/27 DT (EcoRI & XbaI)||3.0||8/30 tRNA-Spinach (EcoRI & SpeI)||12.5||3.5
 +
|-
 +
|experiment||8/20 DT (EcoRI & XbaI)||6.6||8/30 RBS-lysis2 (EcoRI & SpeI)||12.5||3.5
 +
|-
 +
|experiment||8/22 J23100 (SpeI & PstI)||--||RBS-tetR-DT (XbaI & PstI)||--||3.5
 +
|-
 +
|experiment||8/22 Plac (SpeI & PstI)||8.0||8/18 RBS-lacZ&alpha;-DT (XbaI & PstI)||10||3.5
 +
|}
 +
 
 +
Samples were evaporeted used evaporator into about 7 µL
 +
</div>
 +
 
 +
===Miniprep===
 +
<div class="experiment">
 +
<span class="author">Kojima and Nakamoto</span>
 +
{|class="wikitable"
 +
!DNA||concentration[&micro;g/mL]||260/280||260/230
|-
|-
-
|experiment||8/30 pSB1C3(XbaI&Pst)||12.5||8/30 pT181 attenuator(XbaI&PstI)||25||3.5
+
|8/30 Plux-RBS-GFP-DT-(1)||124||1.77||1.87
|-
|-
-
|experiment||8/30 pSB1C3(XbaI&Pst)||12.5||8/30 pT181 antisense(XbaI&PstI)||25||3.5
+
|8/30 Plux-RBS-GFP-DT-(2)||213||1.75||1.52
|-
|-
-
|experiment||8/20 DT||6.6||8/30 pT181 attenuator(2)||12.5||3.5
+
|8/30 Pbad/araC-RBS-RFP||427||1.70||1.56
|-
|-
-
|experiment||8/20 DT||6.6||8/30 pT181 antisense||12.5||3.5
+
|8/30 Pbad/araC-RBS-RFP||670||1.30||0.83
|-
|-
-
|experiment||8/27 DT||3.0||8/30 tRNA-Spinach(EcoRI&SpeI)||12.5||3.5
+
|8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)||332||1.73||2.04
|-
|-
-
|experiment||8/20 DT||6.6||8/30 RBS-lysis2||12.5||3.5
+
|8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)||478||1.74||2.09
|-
|-
-
|experiment||8/22 J23100(SpeI&PstI)||--||RBS-tetR-DT||--
+
|8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)||146||1.77||2.11
|-
|-
-
|experiment||8/22 Plac(SpeI&PstI)||8.0||8/18 RBS-lacZα-DT||10
+
|8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)||454||1.75||2.19
|}
|}
</div>
</div>

Latest revision as of 03:52, 28 September 2013

Contents

Aug 31

Gel Extraction

No name

LaneDNAEnzyme
1100bp ladder--
2----
3RBS-lysis2-(9)EcoRI & SpeI
4
5----
6pSB1C3(1)EcoRI & SpeI
7
8----
9pSB1C3(1)XbaI & PstI
10

Igku Aug30 Gel Extraction(N1).jpg
Igku Aug30 Gel Extraction(N2-1).jpg

LaneDNAEnzyme
1100bp ladder--
2tRNA-SpinachEcoRI & SpeI
3
4----
5pT181 attenuator-(2)EcoRI & SpeI
6
7pT181 attenuator-(2)XbaI & PstI
8
9----
10pT181 antisense-(2)EcoRI & SpeI
11

Igku Aug31 Gel Extraction(N2-2).jpg
Igku Aug31 Gel Extraction(N2-3).jpg

LaneDNAEnzyme
1100bp ladder--
2pT181 antisenseXbaRI & PstI
3
4----
5tetR aptamer 12_1R-(1)EcoRI & SpeI
6

Igku Aug31 Gel Extraction(N3-1).jpg
Igku Aug31 Gel Extraction(N3-2).jpg

Nameconcentration[µg/mL]260/280260/230
RBS-lysis2(EcoRI&SpeI)4.02.690.24
pSB1C3(EcoRI&SpeI)6.02.480.05
pSB1C3(XbaI&PstI)------
tRNA-Spinach------
pT181 attenuator-(2)(EcoRI&SpeI)------
pT181 anttenuator-(2)(SbaI&PstI)------
pT181 antisense-(2)(EcoRI&SpeI)------
pT181 antisense-(2)(XbaI&PstI)------
tetR aptamer 12_1R(EcoRI&SpeI)------

Ligation

Nakamoto and Hirano

stateVectorInserterLigation High ver.2
experiment8/30 pSB1C3 (EcoRI & SpeI)6.68/30 tRNA-Spinach (EcoRI & SpeI)12.53.5
experiment8/30 pSB1C3 (EcoRI & SpeI)6.68/30 pT181 antisense (EcoRI & SpeI)12.53.5
experiment8/30 pSB1C3 (XbaI & Pst)12.58/30 pT181 attenuator (XbaI & PstI)253.5
experiment8/30 pSB1C3 (XbaI & Pst)12.58/30 pT181 antisense (XbaI & PstI)253.5
experiment8/20 DT (EcoRI & XbaI)6.68/30 pT181 attenuator(2) (EcoRI & SpeI)12.53.5
experiment8/20 DT (EcoRI & XbaI)6.68/30 pT181 antisense (EcoRI & SpeI)12.53.5
experiment8/27 DT (EcoRI & XbaI)3.08/30 tRNA-Spinach (EcoRI & SpeI)12.53.5
experiment8/20 DT (EcoRI & XbaI)6.68/30 RBS-lysis2 (EcoRI & SpeI)12.53.5
experiment8/22 J23100 (SpeI & PstI)--RBS-tetR-DT (XbaI & PstI)--3.5
experiment8/22 Plac (SpeI & PstI)8.08/18 RBS-lacZα-DT (XbaI & PstI)103.5

Samples were evaporeted used evaporator into about 7 µL

Miniprep

Kojima and Nakamoto

DNAconcentration[µg/mL]260/280260/230
8/30 Plux-RBS-GFP-DT-(1)1241.771.87
8/30 Plux-RBS-GFP-DT-(2)2131.751.52
8/30 Pbad/araC-RBS-RFP4271.701.56
8/30 Pbad/araC-RBS-RFP6701.300.83
8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(1)3321.732.04
8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(2)4781.742.09
8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(3)1461.772.11
8/30 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT-(4)4541.752.19
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