Template:Kyoto/Notebook/Aug 10

From 2013.igem.org

(Difference between revisions)
(→LB Medium Plate)
(→LB Medium Plate(+Ampicillin))
 
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Line 1: Line 1:
==Aug 10==
==Aug 10==
-
===Liquid Culture===
+
===Colony PCR===
<div class="experiment">
<div class="experiment">
<span class="author">Nakamoto</span>
<span class="author">Nakamoto</span>
{| class="wikitable"
{| class="wikitable"
-
!Sample||medium
+
!Sample
|-
|-
-
|8/9 BBa-J23106||LB(+amp)  
+
|8/9 BBa_J44000 -(1)
|-
|-
-
|8/9 BBa-J23100-1||LB(+amp)  
+
|8/9 BBa_J44000 -(2)
|-
|-
-
|8/9 BBa-J23100-2||LB(+amp)  
+
|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||30 sec||30 cycles
 +
|}
 +
{| class="wikitable"
 +
!Sample
 +
|-
 +
|8/9 BBa_J23106
 +
|-
 +
|8/9 BBa_J23100 -(1)
 +
|-
 +
|8/9 BBa_J23100 -(2)
 +
|-
 +
|NC
 +
|}
 +
{| class="wikitable"
 +
!PreDenature||Denature||Annealing||Extension||cycle
|-
|-
-
|8/9 BBa-J44000-1||LB(+amp)
+
|94 &deg;C||94 &deg;C||55 &deg;C||68 &deg;C||--
|-
|-
-
|8/9 BBa-J44000-2||LB(+amp)
+
|5 min||30 sec||30 sec||1 min||30 cycles
|}
|}
</div>
</div>
-
===Waking Up Parts===
+
===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">
<div class="experiment">
<span class="author">Nakamoto</span>
<span class="author">Nakamoto</span>
{| class="wikitable"
{| class="wikitable"
-
!Sample
+
!Sample||medium
|-
|-
-
|BBa-Q04400(RBS-tetR-DT-Ptet)pSB2K3
+
|8/9 BBa-J23106||LB(+Amp)  
|-
|-
-
|BBa-B0034(RBS)pSB1A2
+
|8/9 BBa-J23100-1||LB(+Amp)  
|-
|-
-
|BBa-I732020(RBS-lacZ&alpha;-DTt)pSB1C3
+
|8/9 BBa-J23100-2||LB(+Amp)  
|-
|-
-
|BBa-P0412(RBS-lacI-DT)pSB1C3
+
|8/9 BBa-J44000-1||LB(+Amp)  
|-
|-
-
|BBa-R0011(Plac)pSB1C3
+
|8/9 BBa-J44000-2||LB(+Amp)  
|}
|}
</div>
</div>
Line 50: Line 102:
|0.4M NaOHaq||500&micro;L
|0.4M NaOHaq||500&micro;L
|-
|-
-
|Agarose Pouder||2g　
+
|Agarose Pouder||2g
|-
|-
|Ampicillin||40&micro;L
|Ampicillin||40&micro;L
Line 57: Line 109:
===Transformation===
===Transformation===
-
===Gel Making===
+
<div class="experiment">
 +
<span class="author">Hirano</span>
 +
{| class="wikitable"
 +
!Name||Sample||Competent Cells||Total||Plate
 +
|-
 +
|tetR||3&micro;L||19&micro;L||22&micro;L||Kan
 +
|-
 +
|RBS||3&micro;L||19&micro;L||22&micro;L||Amp
 +
|-
 +
|lacZ&alpha;||3&micro;L||19&micro;L||22&micro;L||CP
 +
|-
 +
|lacI||3&micro;L||19&micro;L||22&micro;L||CP
 +
|-
 +
|Plac||3&micro;L||19&micro;L||22&micro;L||CP
 +
|}
 +
</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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