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Team:HokkaidoU Japan/Notebook/Promoter RandomPromtoer-B0034-LacZ-B0015
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| - | <h2></h2> | + | <h2>RandomPromtoer-B0034-LacZ-B0015</h2> |
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<div class="step-tile"><h2>Start</h2></div> | <div class="step-tile"><h2>Start</h2></div> | ||
</li> | </li> | ||
| - | + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat May 11</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-lacZα-B0015 on pSB1C3 </div> </div> </div> </li> | |
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Mini-prep</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> K1084007-B0034-lacZα-B0015 on pSB1C3 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR & PCR purification</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> random promoter<br> random-oripro-f, 200b dn, KOD plus Neo </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title"> Digestion & Gel extraction</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> Cut random promoter-B0034-lacZα-B0015 with XbaI, PstI (using 10×M buffer)<br> Cut pSB1C3 with XbaI, PstI (using 10×M buffer) </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">EtOH precipitation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun May 12</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015<br> pSB1C3 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Ligation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon May 13</time> </li> </ul> <div class="step-recipe"> Ligate random promoter-B0034-lacZα-B0015 with pSB1C3 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon May 13</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 on pSB1C3<br> 1µL DNA to DH5α </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue May 14</time> </li> </ul> <div class="step-recipe"> Cultivation time for transformation product was elongated from 18 hrs to 24 hrs.<br> About 10 blue colonies occurred after the cultivation. </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Tue May 14</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 on pSB1C3<br> 5µL DNA to DH5α </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">EtOH precipitation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed May 15</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 on pSB1C3(ligation product) </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed May 15</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 on pSB1C3<br> 1µL DNA to DH5α<br> </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Ligation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri May 17</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 with pSB1C3 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Transformation</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon May 20</time> </li> </ul> <div class="step-recipe"> Each samples were transformed and cultivated at 37°C or 30°C<br> <br> Ligation 30 min, cultivate at 37°C<br> Ligation 30 min, cultivate at 30°C<br> Ligation overnight, cultivate at 37°C<br> Ligation overnight, cultivate at 30°C </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Correction</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu May 23</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 on pSB1C3<br> In 5/20, 5 µL of DNA solution was used for transformation instead of 1 µL of DNA solution for transformation. </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu May 23</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 on pSB1C3 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Colony PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu May 23</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015<br> EX-F, PS-R </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu May 24, Sun May 26, Tue May 28</time> </li> </ul> <div class="step-recipe"> Expression comparison<br> random promoter-B0034-lacZα-B0015 on pSB1C3 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Mini-prep </h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed May 29</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 on pSB1C3 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR for sequencing</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri May 31</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015<br> 100b up, 200b dn, KOD Plus Neo </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">EtOH for sequencing</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri May 31</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Sequencing</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri May 31</time> </li> </ul> <div class="step-recipe"> random promoter-B0034-lacZα-B0015 </div> </div> </div> </li> | ||
<li class="step complete"> | <li class="step complete"> | ||
<div class="step-tile"><h2>Complete!</h2></div> | <div class="step-tile"><h2>Complete!</h2></div> | ||
Latest revision as of 06:58, 27 September 2013
Maestro E.coli
Notebook
RandomPromtoer-B0034-LacZ-B0015
-
Start
-
Culture
K1084007-B0034-lacZα-B0015 on pSB1C3 -
Mini-prep
K1084007-B0034-lacZα-B0015 on pSB1C3 -
PCR & PCR purification
random promoter
random-oripro-f, 200b dn, KOD plus Neo -
Digestion & Gel extraction
Cut random promoter-B0034-lacZα-B0015 with XbaI, PstI (using 10×M buffer)
Cut pSB1C3 with XbaI, PstI (using 10×M buffer) -
EtOH precipitation
random promoter-B0034-lacZα-B0015
pSB1C3 -
Ligation
Ligate random promoter-B0034-lacZα-B0015 with pSB1C3 -
Transformation
random promoter-B0034-lacZα-B0015 on pSB1C3
1µL DNA to DH5α -
Culture
Cultivation time for transformation product was elongated from 18 hrs to 24 hrs.
About 10 blue colonies occurred after the cultivation. -
Transformation
random promoter-B0034-lacZα-B0015 on pSB1C3
5µL DNA to DH5α -
EtOH precipitation
random promoter-B0034-lacZα-B0015 on pSB1C3(ligation product) -
Transformation
random promoter-B0034-lacZα-B0015 on pSB1C3
1µL DNA to DH5α
-
Ligation
random promoter-B0034-lacZα-B0015 with pSB1C3 -
Transformation
Each samples were transformed and cultivated at 37°C or 30°C
Ligation 30 min, cultivate at 37°C
Ligation 30 min, cultivate at 30°C
Ligation overnight, cultivate at 37°C
Ligation overnight, cultivate at 30°C -
Correction
random promoter-B0034-lacZα-B0015 on pSB1C3
In 5/20, 5 µL of DNA solution was used for transformation instead of 1 µL of DNA solution for transformation. -
Culture
random promoter-B0034-lacZα-B0015 on pSB1C3 -
Colony PCR
random promoter-B0034-lacZα-B0015
EX-F, PS-R -
Culture
Expression comparison
random promoter-B0034-lacZα-B0015 on pSB1C3 -
Mini-prep
random promoter-B0034-lacZα-B0015 on pSB1C3 -
PCR for sequencing
random promoter-B0034-lacZα-B0015
100b up, 200b dn, KOD Plus Neo -
EtOH for sequencing
random promoter-B0034-lacZα-B0015 -
Sequencing
random promoter-B0034-lacZα-B0015 -
Complete!
