Template:Kyoto/Notebook/Sep 6

From 2013.igem.org

(Difference between revisions)
(→Miniprep)
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==Sep 6==
==Sep 6==
-
===Liquid Culture===
 
-
<!-- ここから -->
 
-
<div class="experiment">
 
-
<span class="author">No name</span>
 
-
{| class="wikitable"
 
-
!Sample||medium
 
-
|-
 
-
|9/4 RBS-lysis3＋DT3 ||Plusgrow medium(+CP)
 
-
|-
 
-
|9/4 RBS-lysis1＋DT3 ||　Plusgrow medium(+CP)
 
-
|-
 
-
|9/4 psB4K5||Plusgrow medium(+K)
 
-
|}
 
-
</div>
 
-
 
-
===Colony PCR===
 
-
<div class="experiment">
 
-
<span class="author">No name</span>
 
-
{| class="wikitable"
 
-
!|Sample||base 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)||-
 
-
|}
 
-
{| class="wikitable"
 
-
!|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)||-
 
-
|-
 
-
|Plac-PT181-antisense-(3)||-
 
-
|-
 
-
|Plac-PT181-antisense-(4)||-
 
-
|}
 
-
 
-
===Miniprep===
 
-
<div class="experiment">
 
-
<span class="author">Honda and Kojima</span>
 
-
{|class="wikitable"
 
-
!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>
 
-
<!-- ここまでをコピーしてね -->
 
-
 
===Restriction Enzyme Digestion===
===Restriction Enzyme Digestion===
<div class="experiment">
<div class="experiment">
-
<span class="author">Murata, Murakami, Nakamoto, Ashida, Matsumoto and Yoshida</span>
+
<span class="author">Tatsui</span>
{| class="wikitable"
{| class="wikitable"
-
! ||8/11 J23100 -(2)||SpeI||PstI||Buffer||MilliQ||total
+
! ||9/6 RBS-lysis2-DT (78.3 µg/mL)||EcoRI||SpeI||XbaI||PstI||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||4.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||21&micro;L||30&micro;L
+
|2 cuts||22&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||0&micro;L||30&micro;L
|-
|-
-
|1 cut||1.3&micro;L||0.2&micro;L||--||1.0&micro;L||7.5&micro;L||10&micro;L
+
|NC||1.3&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||6.7&micro;L||10&micro;L
-
|-
+
-
|1 cut||1.3&micro;L||--||0.2&micro;L||1.0&micro;L||7.5&micro;L||10&micro;L
+
-
|-
+
-
|NC||1.3&micro;L||--||--||1.0&micro;L||7.7&micro;L||10&micro;L
+
|}
|}
*at 37&deg;C, for 1h
*at 37&deg;C, for 1h
 +
{| class="wikitable"
{| class="wikitable"
-
! ||8/15 B0034(RBS) -(2)||SpeI||PstI||Buffer||MilliQ||total
+
! ||aptamer-DT (251.7 µg/mL)||EcoRI||SpeI||XbaI||PstI||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||6.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||19&micro;L||30&micro;L
+
|2 cuts||0.4&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||3&micro;L||3&micro;L||14.1&micro;L||30&micro;L
|-
|-
-
|1 cut||2.0&micro;L||0.2&micro;L||--||1.0&micro;L||6.8&micro;L||10&micro;L
+
|NC||0.4&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||7.6&micro;L||10&micro;L
-
|-
+
-
|1 cut||2.0&micro;L||--||0.2&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
+
-
|-
+
-
|NC||2.0&micro;L||--||--||1.0&micro;L||7.0&micro;L||10&micro;L
+
|}
|}
*at 37&deg;C, for 1h
*at 37&deg;C, for 1h
 +
{| class="wikitable"
{| class="wikitable"
-
! ||8/17 K117000(lysis1) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
+
! ||Pλ-luxI (86.4 µg/mL)||EcoRI||SpeI||XbaI||PstI||BSA||Buffer||MilliQ||total
|-
|-
-
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L||20&micro;L||30&micro;L
+
|2 cuts||23.1&micro;L||1&micro;L||1&micro;L||0&micro;L||0&micro;L||3&micro;L||3&micro;L||1.9&micro;L||30&micro;L
|-
|-
-
|1 cut||1.0&micro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
+
|NC||1.2&micro;L||0&micro;L||0&micro;L||0&micro;L||0&micro;L||1&micro;L||1&micro;L||6.8&micro;L||10&micro;L
-
|-
+
-
|1 cut||1.0&micro;L||--||0.2&micro;L||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
+
-
|-
+
-
|NC||1.0&micro;L||--||--||1.0&micro;L||1.0&micro;L||7.0&micro;L||10&micro;L
+
|}
|}
*at 37&deg;C, for 1h
*at 37&deg;C, for 1h
-
{| class="wikitable"
 
-
! ||8/17 K748002(lysis3) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
 
-
|-
 
-
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L||20&micro;L||30&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||--||0.2&micro;L||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|NC||1.0&micro;L||--||--||1.0&micro;L||1.0&micro;L||7.0&micro;L||10&micro;L
 
-
|}
 
-
*at 37&deg;C, for 1h
 
-
{| class="wikitable"
 
-
! ||8/17 I13504(RBS-GFP-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
 
-
|-
 
-
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L||20&micro;L||30&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||--||0.2&micro;L||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|NC||1.0&micro;L||--||--||1.0&micro;L||1.0&micro;L||7.0&micro;L||10&micro;L
 
-
|}
 
-
*at 37&deg;C, for 1h
 
-
{| class="wikitable"
 
-
! ||8/17 I0462(RBS-luxR-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
 
-
|-
 
-
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L||20&micro;L||30&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||--||0.2&micro;L||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|NC||1.0&micro;L||--||--||1.0&micro;L||1.0&micro;L||7.0&micro;L||10&micro;L
 
-
|}
 
-
*at 37&deg;C, for 1h
 
-
{| class="wikitable"
 
-
! ||8/17 I732020(RBS-lacZ&alpha;-DT) -(1)||XbaI||PstI||BSA||Buffer||MilliQ||total
 
-
|-
 
-
|2 cuts||2.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||3.0&micro;L||20&micro;L||30&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||0.2&micro;L||--||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|1 cut||1.0&micro;L||--||0.2&micro;L||1.0&micro;L||1.0&micro;L||6.8&micro;L||10&micro;L
 
-
|-
 
-
|NC||1.0&micro;L||--||--||1.0&micro;L||1.0&micro;L||7.0&micro;L||10&micro;L
 
-
|}
 
-
*at 37&deg;C, for 1h
 
-
{| class="wikitable"
 
-
! ||8/11 R6061 -(1)||SpeI||PstI||Buffer||MilliQ||total
 
-
|-
 
-
|2 cuts||8.0&micro;L||1.0&micro;L||1.0&micro;L||3.0&micro;L||17&micro;L||30&micro;L
 
-
|-
 
-
|1 cut||3.0&micro;L||0.2&micro;L||--||1.0&micro;L||5.8&micro;L||10&micro;L
 
-
|-
 
-
|1 cut||3.0&micro;L||--||0.2&micro;L||1.0&micro;L||5.8&micro;L||10&micro;L
 
-
|-
 
-
|NC||3.0&micro;L||--||--||1.0&micro;L||6.0&micro;L||10&micro;L
 
-
|}
 
-
*at 37&deg;C, for 1h
 
-
</div>
 
-
 
-
===Gel Extraction===
 
-
<div class="experiment">
 
-
<span class="author">Ashida</span>
 
-
{| class="wikitable"
 
-
!Lane||DNA||Enzyme
 
-
|-
 
-
|1||100bp ladder||--
 
-
|-
 
-
|3||8/18 K117000(lysis2) -1||XbaI & PstI
 
-
|-
 
-
|5||8/18 K117000(lysis2) -1||XbaI & PstI
 
-
|}
 
-
[[File:igku_Aug19electrophoresis3]]<br>
 
-
[[File:igku_Aug19electrophoresis4]]<br>
 
-
{| class="wikitable"
 
-
!Name||concentration[&micro;g/mL]||260/280||260/230
 
-
|-
 
-
|8/18 lysis2-1(XbaI & PstI)||3.1||3.64||0.19
 
-
|-
 
-
|8/18 lysis2-1(XbaI & PstI)||3.4||1.89||0.13
 
-
|}
 
-
 
-
</div>
 
-
 
-
 
-
===Liquid Culture===
 
-
<div class="experiment">
 
-
<span class="author">Nakamoto</span>
 
-
{| class="wikitable"
 
-
!Sample||medium
 
-
|-
 
-
|8/15 BBa_K112806(lysis2) -(1)||Plusgrow medium(+CP)
 
-
|-
 
-
|8/15 BBa_Q04400(tetR) -(1)||Plusgrow medium(+Amp)
 
-
|-
 
-
|}
 
-
</div>
 
-
 
-
 
-
===Colony PCR===
 
-
<div class="experiment">
 
-
<span class="author">Nakamoto</span>
 
-
{| class="wikitable"
 
-
!|Sample||base pair
 
-
|-
 
-
|8/15 BBa_K112806(lysis2) -(1)||828
 
-
|-
 
-
|negative control||--
 
-
|}
 
-
{| class="wikitable"
 
-
!PreDenature||Denature||Annealing||Extension||cycle
 
-
|-
 
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 
-
|-
 
-
|5min||30s||30s||1min||30cycles
 
-
|}
 
-
{| class="wikitable"
 
-
!Sample||base pair
 
-
|-
 
-
|8/10 BBa_Q0400 -(1)||3995
 
-
|-
 
-
|negative control||--
 
-
|}
 
-
{| class="wikitable"
 
-
!PreDenature||Denature||Annealing||Extension||cycle
 
-
|-
 
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 
-
|-
 
-
|5min||30s||30s||30s||30cycles
 
-
|}
 
-
 
-
===Colony PCR===
 
-
<div class="experiment">
 
-
<span class="author">Nakamoto</span>
 
-
{| class="wikitable"
 
-
!|Sample||base pair
 
-
|-
 
-
|8/15 BBa_K112806(lysis2) -(1)||828
 
-
|-
 
-
|negative control||--
 
-
|}
 
-
{| class="wikitable"
 
-
!PreDenature||Denature||Annealing||Extension||cycle
 
-
|-
 
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 
-
|-
 
-
|5min||30s||30s||1min||30cycles
 
-
|}
 
-
{| class="wikitable"
 
-
!Sample||base pair
 
-
|-
 
-
|8/10 BBa_Q0400 -(1)||3995
 
-
|-
 
-
|negative control||--
 
-
|}
 
-
{| class="wikitable"
 
-
!PreDenature||Denature||Annealing||Extension||cycle
 
-
|-
 
-
|94&deg;C||94&deg;C||55&deg;C||68&deg;C||--
 
-
|-
 
-
|5min||30s||30s||30s||30cycles
 
-
|}
 

Revision as of 09:17, 24 September 2013

Sep 6

Restriction Enzyme Digestion

Tatsui

9/6 RBS-lysis2-DT (78.3 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts22µL0µL0µL1µL1µL3µL3µL0µL30µL
NC1.3µL0µL0µL0µL0µL1µL1µL6.7µL10µL
  • at 37°C, for 1h
aptamer-DT (251.7 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts0.4µL0µL0µL1µL1µL3µL3µL14.1µL30µL
NC0.4µL0µL0µL0µL0µL1µL1µL7.6µL10µL
  • at 37°C, for 1h
Pλ-luxI (86.4 µg/mL)EcoRISpeIXbaIPstIBSABufferMilliQtotal
2 cuts23.1µL1µL1µL0µL0µL3µL3µL1.9µL30µL
NC1.2µL0µL0µL0µL0µL1µL1µL6.8µL10µL
  • at 37°C, for 1h
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