Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Sep 6)
(→Colony PCR)
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===Colony PCR===
===Colony PCR===
<div class="experiment">
<div class="experiment">
-
<span class="author">Nakamoto</span>
+
<span class="author">No name</span>
{| class="wikitable"
{| class="wikitable"
!|Sample||base pair
!|Sample||base pair
|-
|-
-
|8/15 BBa_K112806(lysis2) -(1)||828
+
|Pconst-spinach-DT-(1)||-
|-
|-
-
|negative control||--
+
|Pconst-spinach-DT-(2)||-
-
|}
+
-
{| class="wikitable"
+
-
!PreDenature||Denature||Annealing||Extension||cycle
+
|-
|-
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
+
|Pconst-spinach-DT-(3)||-
|-
|-
-
|5min||30s||30s||1min||30cycles
+
|Pconst-spinach-DT-(4)||-
-
|}
+
-
{| class="wikitable"
+
-
!Sample||base pair
+
|-
|-
-
|8/10 BBa_Q0400 -(1)||3995
+
|Plac-spinach-DT-(1)||-
|-
|-
-
|negative control||--
+
|Plac-spinach-DT-(2)||-
 +
|-
 +
|Plac-spinach-DT-(3)||-
 +
|-
 +
|Plac-spinach-DT-(4)||-
 +
|-
 +
|Pcon-PT181-antisense-(1)||-
 +
|-
 +
|Pcon-PT181-antisense-(2)||-
 +
|-
 +
|Pcon-PT181-antisense-(3)||-
 +
|-
 +
|Pcon-PT181-antisense-(4)||-
|}
|}
{| class="wikitable"
{| class="wikitable"
-
!PreDenature||Denature||Annealing||Extension||cycle
+
!|Sample||base pair
 +
|-
 +
|Ptet-PT181-antisense-(1)||-
 +
|-
 +
|Ptet-PT181-antisense-(2)||-
 +
|-
 +
|Ptet-PT181-antisense-(3)||-
 +
|-
 +
|Ptet-PT181-antisense-(4)||-
 +
|-
 +
|Plac-PT181-antisense-(1)||-
 +
|-
 +
|Plac-PT181-antisense-(2)||-
|-
|-
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
+
|Plac-PT181-antisense-(3)||-
|-
|-
-
|5min||30s||30s||30s||30cycles
+
|Plac-PT181-antisense-(4)||-
|}
|}

Revision as of 07:51, 24 September 2013

Contents

Sep 6

Liquid Culture

No name

Samplemedium
9/4 RBS-lysis3＋DT3 Plusgrow medium(+CP)
9/4 RBS-lysis1＋DT3 　Plusgrow medium(+CP)
9/4 psB4K5Plusgrow medium(+K)

Colony PCR

No name

Samplebase pair
Pconst-spinach-DT-(1)-
Pconst-spinach-DT-(2)-
Pconst-spinach-DT-(3)-
Pconst-spinach-DT-(4)-
Plac-spinach-DT-(1)-
Plac-spinach-DT-(2)-
Plac-spinach-DT-(3)-
Plac-spinach-DT-(4)-
Pcon-PT181-antisense-(1)-
Pcon-PT181-antisense-(2)-
Pcon-PT181-antisense-(3)-
Pcon-PT181-antisense-(4)-
Samplebase pair
Ptet-PT181-antisense-(1)-
Ptet-PT181-antisense-(2)-
Ptet-PT181-antisense-(3)-
Ptet-PT181-antisense-(4)-
Plac-PT181-antisense-(1)-
Plac-PT181-antisense-(2)-
Plac-PT181-antisense-(3)-
Plac-PT181-antisense-(4)-

Miniprep

No name

DNAconcentration[µg/mL]260/280260/230
(・▽・)　　

Restriction Enzyme Digestion

Murata, Murakami, Nakamoto, Ashida, Matsumoto and Yoshida

8/11 J23100 -(2)SpeIPstIBufferMilliQtotal
2 cuts4.0µL1.0µL1.0µL3.0µL21µL30µL
1 cut1.3µL0.2µL--1.0µL7.5µL10µL
1 cut1.3µL--0.2µL1.0µL7.5µL10µL
NC1.3µL----1.0µL7.7µL10µL
  • at 37°C, for 1h
8/15 B0034(RBS) -(2)SpeIPstIBufferMilliQtotal
2 cuts6.0µL1.0µL1.0µL3.0µL19µL30µL
1 cut2.0µL0.2µL--1.0µL6.8µL10µL
1 cut2.0µL--0.2µL1.0µL6.8µL10µL
NC2.0µL----1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 K117000(lysis1) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 K748002(lysis3) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 I13504(RBS-GFP-DT) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 I0462(RBS-luxR-DT) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/17 I732020(RBS-lacZα-DT) -(1)XbaIPstIBSABufferMilliQtotal
2 cuts2.0µL1.0µL1.0µL3.0µL3.0µL20µL30µL
1 cut1.0µL0.2µL--1.0µL1.0µL6.8µL10µL
1 cut1.0µL--0.2µL1.0µL1.0µL6.8µL10µL
NC1.0µL----1.0µL1.0µL7.0µL10µL
  • at 37°C, for 1h
8/11 R6061 -(1)SpeIPstIBufferMilliQtotal
2 cuts8.0µL1.0µL1.0µL3.0µL17µL30µL
1 cut3.0µL0.2µL--1.0µL5.8µL10µL
1 cut3.0µL--0.2µL1.0µL5.8µL10µL
NC3.0µL----1.0µL6.0µL10µL
  • at 37°C, for 1h

Gel Extraction

Ashida

LaneDNAEnzyme
1100bp ladder--
38/18 K117000(lysis2) -1XbaI & PstI
58/18 K117000(lysis2) -1XbaI & PstI

File:Igku Aug19electrophoresis3
File:Igku Aug19electrophoresis4

Nameconcentration[µg/mL]260/280260/230
8/18 lysis2-1(XbaI & PstI)3.13.640.19
8/18 lysis2-1(XbaI & PstI)3.41.890.13


Liquid Culture

Nakamoto

Samplemedium
8/15 BBa_K112806(lysis2) -(1)Plusgrow medium(+CP)
8/15 BBa_Q04400(tetR) -(1)Plusgrow medium(+Amp)


Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles
Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles

Colony PCR

Nakamoto

Samplebase pair
8/15 BBa_K112806(lysis2) -(1)828
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s1min30cycles
Samplebase pair
8/10 BBa_Q0400 -(1)3995
negative control--
PreDenatureDenatureAnnealingExtensioncycle
94°C94°C55°C68°C--
5min30s30s30s30cycles
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