Template:Kyoto/Notebook/Aug 24

From 2013.igem.org

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Line 39: Line 39:
|8/22 pT181 antisense (EcoRI & SpeI)||8||1.13||0.82
|8/22 pT181 antisense (EcoRI & SpeI)||8||1.13||0.82
|-
|-
-
|8/22 Fusion3m2 attenuator (XbaI & PstI)||5||2.277||1.50
+
|8/22 Fusion3m2 attenuator (XbaI & PstI)||5||2.27||1.50
|-
|-
|8/22 Fusion3m2 attenuator (EcoRI & SpeI)||34||1.16||1.02
|8/22 Fusion3m2 attenuator (EcoRI & SpeI)||34||1.16||1.02
Line 53: Line 53:
===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
-
<!-- こっから -->
 
<div class="experiment">
<div class="experiment">
<span class="author">No name</span>
<span class="author">No name</span>
{| class="wikitable"
{| class="wikitable"
-
!tetR aptamer 12_P(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total
+
! ||tetR aptamer 12_P(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total
|-
|-
-
|2.7&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||23&micro;L||30&micro;L
+
|2 cuts||2.7&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||23&micro;L||30&micro;L
|-
|-
-
|0.3&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.6&micro;L||10&micro;L
+
|NC||0.3&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.6&micro;L||10&micro;L
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!tetR aptamer 12_1R(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total
+
! ||tetR aptamer 12_1R(2)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total
|-
|-
-
|2.6&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||23.1&micro;L||30&micro;L
+
|2 cuts||2.6&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||23.1&micro;L||30&micro;L
|-
|-
-
|0.3&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.6&micro;L||10&micro;L
+
|NC||0.3&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.6&micro;L||10&micro;L
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!tetR aptamer 12_M(1)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total
+
! ||tetR aptamer 12_M(1)||EcoRI||SpeI||10x Buffer B||100x BSA||MilliQ||total
|-
|-
-
|2.7&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||23&micro;L||30&micro;L
+
|2 cuts||2.7&micro;L||0.5&micro;L||0.5&micro;L||3&micro;L||0.3&micro;L||23&micro;L||30&micro;L
|-
|-
-
|0.3&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.6&micro;L||10&micro;L
+
|NC||0.3&micro;L||0&micro;L||0&micro;L||1&micro;L||0.1&micro;L||8.6&micro;L||10&micro;L
|}
|}
 +
</div>
===Master Plate===
===Master Plate===
Line 99: Line 99:
===Electrophoresis===
===Electrophoresis===
 +
<div class="experiment">
 +
<span class="author">No name</span>
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||-||-
 +
|-
 +
|2||tetR aptamer 12_P||EcoRI||SpeI
 +
|-
 +
|3||tetR aptamer 12_P||-||-
 +
|-
 +
|4||tetR aptamer 12_1R||EcoRI||SpeI
 +
|-
 +
|5||tetR aptamer 12_1R||-||-
 +
|-
 +
|6||tetR aptamer 12_1M||EcoRI||SpeI
 +
|-
 +
|7||tetR aptamer 12_1M||-||-
 +
|-
 +
|8||100bp ladder||-||-
 +
|}<br>
 +
[[File:Igku Aug24 Electrophoresis(N1)-pic.jpg]]<br>
 +
<br>
 +
 +
{| class="wikitable"
 +
!Lane||Sample||Enzyme1||Enzyme2
 +
|-
 +
|1||100bp ladder||-||-
 +
|-
 +
|2||tetR aptamer 12_P||EcoRI||SpeI
 +
|-
 +
|3||tetR aptamer 12_1R||EcoRI||SpeI
 +
|-
 +
|4||tetR aptamer 12_1M||EcoRI||SpeI
 +
|-
 +
|5||100bp ladder||-||-
 +
|}
 +
<br>
 +
[[File:Igku Aug24 Electrophoresis(N2)-pic2.jpg]]<br>
 +
 +
<br>
 +
</div>

Latest revision as of 16:18, 25 September 2013

Contents

Aug 24

Gel Extraction

Ashida

Nameconcentration[µg/mL]260/280260/230
8/22 aptamer 12_P (EcoRI & SpeI)8821.3--
8/22 aptamer 12_1R (EcoRI & SpeI)6323.1--
8/22 aptamer 12_1M (EcoRI & SpeI)4022.0--
8/23 Fusion6 antisense (EcoRI & SpeI)6.01.600.17
8/23 Fusion6 antisense (XbaI & PstI)6.01.710.25
8/23 pT181 antisense (EcoRI & SpeI)6.02.220.15
8/23 pT181 antisense (XbaI & PstI)6.02.100.04
8/22 Plac (SpeI & PstI)6.01.470.09
8/22 Fusion1 antisense (EcoRI & SpeI)102.160.01
8/22 J23100 (SpeI & PstI)6.01.520.35
Nameconcentration[µg/mL]260/280260/230
8/22 Spinach (EcoRI & SpeI)8.01.150.77
8/22 Fusion6 antisense (XbaI & PstI)441.070.96
8/22 Fusion1 antisense (XbaI & PstI)2--1.00
8/22 pT181 antisense (XbaI & PstI)3.2.010.40
8/22 pT181 antisense (EcoRI & SpeI)81.130.82
8/22 Fusion3m2 attenuator (XbaI & PstI)52.271.50
8/22 Fusion3m2 attenuator (EcoRI & SpeI)341.161.02
8/22 Fusion1 attenuator (XbaI & PstI)141.400.62
8/22 Fusion1 attenuator (EcoRI & SpeI)161.270.61
8/22 pT181 attenuator (XbaI & PstI)5--0.88
  • These samples were performed ethanol ethanl precipitation.

Restriction Enzyme Digestion

No name

tetR aptamer 12_P(2)EcoRISpeI10x Buffer B100x BSAMilliQtotal
2 cuts2.7µL0.5µL0.5µL3µL0.3µL23µL30µL
NC0.3µL0µL0µL1µL0.1µL8.6µL10µL
tetR aptamer 12_1R(2)EcoRISpeI10x Buffer B100x BSAMilliQtotal
2 cuts2.6µL0.5µL0.5µL3µL0.3µL23.1µL30µL
NC0.3µL0µL0µL1µL0.1µL8.6µL10µL
tetR aptamer 12_M(1)EcoRISpeI10x Buffer B100x BSAMilliQtotal
2 cuts2.7µL0.5µL0.5µL3µL0.3µL23µL30µL
NC0.3µL0µL0µL1µL0.1µL8.6µL10µL

Master Plate

No name

NumberUse LB plate(+Kan)
1entA- #1
2entA- #1
3entA- #1
4entA- #2
5entA- #2
6entA- #2

Electrophoresis

No name

LaneSampleEnzyme1Enzyme2
1100bp ladder--
2tetR aptamer 12_PEcoRISpeI
3tetR aptamer 12_P--
4tetR aptamer 12_1REcoRISpeI
5tetR aptamer 12_1R--
6tetR aptamer 12_1MEcoRISpeI
7tetR aptamer 12_1M--
8100bp ladder--

Igku Aug24 Electrophoresis(N1)-pic.jpg

LaneSampleEnzyme1Enzyme2
1100bp ladder--
2tetR aptamer 12_PEcoRISpeI
3tetR aptamer 12_1REcoRISpeI
4tetR aptamer 12_1MEcoRISpeI
5100bp ladder--


Igku Aug24 Electrophoresis(N2)-pic2.jpg


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