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Team:HokkaidoU Japan/Notebook/RBS SD2,4,6-1C3-Replacement
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| - | <h2></h2> | + | <h2>SD2, 4, 6-1C3 Replacement</h2> |
<div class="workflow-container"> | <div class="workflow-container"> | ||
<ul class="steps"> | <ul class="steps"> | ||
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</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>Wed Jul 17</time> </li> </ul> <div class="step-recipe"> 1C3 vector </div> </div> </div> </li> | |
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Thu Jul 18</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 100up 200dn<br> faild </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>Thu Jul 18</time> </li> </ul> <div class="step-recipe"> 1C3 vector (7/17 cultured) </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 22</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 100up 200dn<br> faild </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 29</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 100up 200dn<br> faild </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Mon Jul 29</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 100up 200dn<br> SD2 faild </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Jul 31</time> </li> </ul> <div class="step-recipe"> R0040-SD2-LacZα-B0015 100up 200dn </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Jul 31</time> </li> </ul> <div class="step-recipe"> Cut R0040-SD(2,4,6)-LacZα-B0015 EcoRI and PstI (using 10×H buffer) </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>Sat Aug 3</time> </li> </ul> <div class="step-recipe"> Ligate R0040-SD(2,4,6)-LacZα-B0015 with mini-preped 1C3 vector </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>Sat Aug 3</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3<br> each of 1µL DNA to DH5α<br> faild<br> </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 Aug 7</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3<br> each of 1µL DNA to DH5α<br> faild </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Aug 9</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 100up 200dn </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sat Aug 10</time> </li> </ul> <div class="step-recipe"> Cut R0040-SD(2,4,6)-LacZα-B0015 EcoRI and PstI (using 10×H buffer) </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Antarctic phophatase</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Aug 11</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 </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>Sun Aug 11</time> </li> </ul> <div class="step-recipe"> Ligate R0040-SD(2,4,6)-LacZα-B0015 with pSB1C3 vector </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 Aug 14</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3<br> each of 1µL DNA to DH5α </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 Aug 15</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-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>Fri Aug 16</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3 </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>Mon Aug 19</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3<br> 100up 200dn<br> faild </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>Thu Aug 22</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3<br> 100up 200dn<br> faild </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">PCR</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Fri Aug 23</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 100up 200dn </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Digestion</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Aug 25</time> </li> </ul> <div class="step-recipe"> Cut R0040-SD(2,4,6)-LacZα-B001 with EcoRI and PstI (using 10×H buffer) </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Antarctic phophatase</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Sun Aug 25</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015<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>Sun Aug 25</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 with pSB1C3 vector </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>Fri Aug 30</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3<br> each of 1µL DNA to DH5α </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 Sep 6</time> </li> </ul> <div class="step-recipe"> R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3<br> 100up 200dn </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 16:39, 27 September 2013
Maestro E.coli
Notebook
SD2, 4, 6-1C3 Replacement
-
Start
-
Culture
1C3 vector -
PCR
R0040-SD(2,4,6)-LacZα-B0015 100up 200dn
faild -
Mini-prep
1C3 vector (7/17 cultured) -
PCR
R0040-SD(2,4,6)-LacZα-B0015 100up 200dn
faild -
PCR
R0040-SD(2,4,6)-LacZα-B0015 100up 200dn
faild -
PCR
R0040-SD(2,4,6)-LacZα-B0015 100up 200dn
SD2 faild -
PCR
R0040-SD2-LacZα-B0015 100up 200dn -
Digestion
Cut R0040-SD(2,4,6)-LacZα-B0015 EcoRI and PstI (using 10×H buffer) -
Ligation
Ligate R0040-SD(2,4,6)-LacZα-B0015 with mini-preped 1C3 vector -
Transformation
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3
each of 1µL DNA to DH5α
faild
-
Transformation
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3
each of 1µL DNA to DH5α
faild -
PCR
R0040-SD(2,4,6)-LacZα-B0015 100up 200dn -
Digestion
Cut R0040-SD(2,4,6)-LacZα-B0015 EcoRI and PstI (using 10×H buffer) -
Antarctic phophatase
R0040-SD(2,4,6)-LacZα-B0015 -
Ligation
Ligate R0040-SD(2,4,6)-LacZα-B0015 with pSB1C3 vector -
Transformation
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3
each of 1µL DNA to DH5α -
Colony PCR
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3 -
Mini-prep
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3 -
Sequencing
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3
100up 200dn
faild -
Sequencing
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3
100up 200dn
faild -
PCR
R0040-SD(2,4,6)-LacZα-B0015 100up 200dn -
Digestion
Cut R0040-SD(2,4,6)-LacZα-B001 with EcoRI and PstI (using 10×H buffer) -
Antarctic phophatase
R0040-SD(2,4,6)-LacZα-B0015
-
Ligation
R0040-SD(2,4,6)-LacZα-B0015 with pSB1C3 vector -
Transformation
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3
each of 1µL DNA to DH5α -
Sequencing
R0040-SD(2,4,6)-LacZα-B0015 on pSB1C3
100up 200dn -
Complete!
