Template:Kyoto/Notebook/Aug 10

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==Aug 10==
==Aug 10==
-
===Ligation===
+
===Colony PCR===
<div class="experiment">
<div class="experiment">
-
<span class="author">Kojima</span>
+
<span class="author">Nakamoto</span>
{| class="wikitable"
{| class="wikitable"
-
!state||colspan="2"|Vector||colspan="2"|Inserter||Ligation High ver.2
+
!Sample
|-
|-
-
|experiment||name||xx &micro;g/mL||name||xx &micro;g/mL||xx &micro;g/mL
+
|8/9 BBa_J44000 -(1)
|-
|-
-
|experiment||8/30 pSB1C3 (EcoRI & SpeI)||6.6||8/30 tRNA-Spinach (EcoRI & SpeI)||12.5||3.5
+
|8/9 BBa_J44000 -(2)
|-
|-
-
|experiment||8/30 pSB1C3 (EcoRI & SpeI)||6.6||8/30 pT181 antisense (EcoRI & SpeI)||12.5||3.5
+
|NC
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
-
|experiment||8/30 pSB1C3 (XbaI & Pst)||12.5||8/30 pT181 attenuator (XbaI & PstI)||25||3.5
+
|94 &deg;C||94 &deg;C||55 &deg;C||68 &deg;C||--
|-
|-
-
|experiment||8/30 pSB1C3 (XbaI & Pst)||12.5||8/30 pT181 antisense (XbaI & PstI)||25||3.5
+
|5 min||30 sec||30 sec||30 sec||30 cycles
 +
|}
 +
{| class="wikitable"
 +
!Sample
|-
|-
-
|experiment||8/20 DT (EcoRI & XbaI)||6.6||8/30 pT181 attenuator(2) (EcoRI & SpeI)||12.5||3.5
+
|8/9 BBa_J23106
 +
|-
 +
|8/9 BBa_J23100 -(1)
 +
|-
 +
|8/9 BBa_J23100 -(2)
 +
|-
 +
|NC
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
 +
|-
 +
|94 &deg;C||94 &deg;C||55 &deg;C||68 &deg;C||--
 +
|-
 +
|5 min||30 sec||30 sec||1 min||30 cycles
 +
|}
 +
</div>
 +
 
 +
===Electrophoresis===
 +
<div class="experiment">
 +
<span class="author">Tatsui and Nakamoto</span>
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
| ||100bp ladder
 +
|-
 +
|4||8/9 BBa_J44000 -1
 +
|-
 +
|5||8/9 BBa_J44000 -2
 +
|-
 +
|6||NC
 +
|}
 +
[[File:Igku Aug10 Electrophoresis BBa 1.jpg]]<br>
 +
{| class="wikitable"
 +
!Lane||Sample
 +
|-
 +
| ||1kbp ladder
 +
|-
 +
|1||8/9 BBa_J23106
 +
|-
 +
|2||8/9 BBa_J23100 -1
 +
|-
 +
|3||8/9 BBa_J23100 -2
 +
|-
 +
|4||NC
 +
|}
 +
[[File:Igku Aug10electrophoresis1.jpg]]<br>
 +
</div>
 +
 
 +
===Liquid Culture===
 +
<div class="experiment">
 +
<span class="author">Nakamoto</span>
 +
{| class="wikitable"
 +
!Sample||medium
 +
|-
 +
|8/9 BBa-J23106||LB(+Amp)
 +
|-
 +
|8/9 BBa-J23100-1||LB(+Amp)
 +
|-
 +
|8/9 BBa-J23100-2||LB(+Amp)  
 +
|-
 +
|8/9 BBa-J44000-1||LB(+Amp)  
 +
|-
 +
|8/9 BBa-J44000-2||LB(+Amp)
 +
|}
 +
</div>
 +
 
 +
===LB Medium Plate(+Ampicillin)===
 +
<div class="experiment">
 +
<span class="author">Nakamoto & Tatsui</span>
 +
{| class="wikitable"
 +
!volume||200ml
 +
|-
 +
|Bacto T2ypton || 2g
 +
|-
 +
|Bacto yeast extract ||1g
 +
|-
 +
|NaCl||1g
 +
|-
 +
|0.4M NaOHaq||500&micro;L
 +
|-
 +
|Agarose Pouder||2g
 +
|-
 +
|Ampicillin||40&micro;L
 +
|}
 +
</div>
 +
 
 +
===Transformation===
 +
<div class="experiment">
 +
<span class="author">Hirano</span>
 +
{| class="wikitable"
 +
!Name||Sample||Competent Cells||Total||Plate
|-
|-
-
|experiment||8/20 DT (EcoRI & XbaI)||6.6||8/30 pT181 antisense (EcoRI & SpeI)||12.5||3.5
+
|tetR||3&micro;L||19&micro;L||22&micro;L||Kan
|-
|-
-
|experiment||8/27 DT (EcoRI & XbaI)||3.0||8/30 tRNA-Spinach (EcoRI & SpeI)||12.5||3.5
+
|RBS||3&micro;L||19&micro;L||22&micro;L||Amp
|-
|-
-
|experiment||8/20 DT (EcoRI & XbaI)||6.6||8/30 RBS-lysis2 (EcoRI & SpeI)||12.5||3.5
+
|lacZ&alpha;||3&micro;L||19&micro;L||22&micro;L||CP
|-
|-
-
|experiment||8/22 J23100 (SpeI & PstI)||--||RBS-tetR-DT (XbaI & PstI)||--||3.5
+
|lacI||3&micro;L||19&micro;L||22&micro;L||CP
|-
|-
-
|experiment||8/22 Plac (SpeI & PstI)||8.0||8/18 RBS-lacZ&alpha;-DT (XbaI & PstI)||10||3.5
+
|Plac||3&micro;L||19&micro;L||22&micro;L||CP
-
|NC||name||xx &micro;g/mL||name||xx &micro;g/mL||xx &micro;g/mL
+
|}
|}
</div>
</div>

Latest revision as of 03:40, 28 September 2013

Contents

Aug 10

Colony PCR

Nakamoto

Sample
8/9 BBa_J44000 -(1)
8/9 BBa_J44000 -(2)
NC
PreDenatureDenatureAnnealingExtensioncycle
94 °C94 °C55 °C68 °C--
5 min30 sec30 sec30 sec30 cycles
Sample
8/9 BBa_J23106
8/9 BBa_J23100 -(1)
8/9 BBa_J23100 -(2)
NC
PreDenatureDenatureAnnealingExtensioncycle
94 °C94 °C55 °C68 °C--
5 min30 sec30 sec1 min30 cycles

Electrophoresis

Tatsui and Nakamoto

LaneSample
100bp ladder
48/9 BBa_J44000 -1
58/9 BBa_J44000 -2
6NC

Igku Aug10 Electrophoresis BBa 1.jpg

LaneSample
1kbp ladder
18/9 BBa_J23106
28/9 BBa_J23100 -1
38/9 BBa_J23100 -2
4NC

Igku Aug10electrophoresis1.jpg

Liquid Culture

Nakamoto

Samplemedium
8/9 BBa-J23106LB(+Amp)
8/9 BBa-J23100-1LB(+Amp)
8/9 BBa-J23100-2LB(+Amp)
8/9 BBa-J44000-1LB(+Amp)
8/9 BBa-J44000-2LB(+Amp)

LB Medium Plate(+Ampicillin)

Nakamoto & Tatsui

volume200ml
Bacto T2ypton 2g
Bacto yeast extract 1g
NaCl1g
0.4M NaOHaq500µL
Agarose Pouder2g
Ampicillin40µL

Transformation

Hirano

NameSampleCompetent CellsTotalPlate
tetR3µL19µL22µLKan
RBS3µL19µL22µLAmp
lacZα3µL19µL22µLCP
lacI3µL19µL22µLCP
Plac3µL19µL22µLCP
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