Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(Colony PCR)
(Miniprep)
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===Miniprep===
===Miniprep===
<div class="experiment">
<div class="experiment">
-
<span class="author">No name</span>
+
<span class="author">Honda and Kojima</span>
{|class="wikitable"  
{|class="wikitable"  
!DNA||concentration[&micro;g/mL]||260/280||260/230  
!DNA||concentration[&micro;g/mL]||260/280||260/230  
|-
|-
-
|(・▽・)|| || ||
+
|PT181 antisense-(1)||238.7 ||1.68 ||1.44
 +
|-
 +
|PT181 antisense-(2)||248.8 ||1.83 ||1.77
 +
|-
 +
|PT181 antisense-(3)||223.2 ||1.78 ||1.61
 +
|-
 +
|PT181 antisense-(4)||220.8 ||1.89 ||1.92
 +
|-
 +
|tetR aptamer-(1)||252.5 ||1.89 ||1.90
 +
|-
 +
|tetR aptamer-(2)||211.2 ||1.90 ||1.99
 +
|-
 +
|PT181 attenuater||102.2 ||1.98 ||2.17
 +
|-
 +
|121R aptamer-DT||251.7 ||1.92 ||2.01
 +
|-
 +
|PSB4K5||212.9 ||1.80 ||1.90
 +
|-
 +
|RBS-lacZα-DT||185.7 ||1.78 ||1.79
 +
|-
 +
|RBS-lysis2-DT-(1)||84.8 ||1.85 ||2.24
|}
|}
</div>
</div>

Revision as of 08:02, 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

Honda and Kojima

DNAconcentration[µg/mL]260/280260/230
PT181 antisense-(1)238.7 1.68 1.44
PT181 antisense-(2)248.8 1.83 1.77
PT181 antisense-(3)223.2 1.78 1.61
PT181 antisense-(4)220.8 1.89 1.92
tetR aptamer-(1)252.5 1.89 1.90
tetR aptamer-(2)211.2 1.90 1.99
PT181 attenuater102.2 1.98 2.17
121R aptamer-DT251.7 1.92 2.01
PSB4K5212.9 1.80 1.90
RBS-lacZα-DT185.7 1.78 1.79
RBS-lysis2-DT-(1)84.8 1.85 2.24

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