"
Team:HokkaidoU Japan/Notebook/Color R0040-B0034-(K592010, K592012, K864401)-B0015
From 2013.igem.org
(Difference between revisions)
| Line 19: | Line 19: | ||
</li> | </li> | ||
| - | + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Get chromoprotein DNA</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"> Great thanks to iGEM Uppsala! </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 Aug 20</time> </li> </ul> <div class="step-recipe"> K592010, K592012, K864401<br> each of 1µL DNA to DH5α </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Colony PCR & Culture</h2> </div> <div class="step-detail"> <div class="step-detail-body"> <ul class="step-detail-trials"> <li> <time>Wed Aug 21</time> </li> </ul> <div class="step-recipe"> K592010, K592012, K864401, B0015<br> We confirmed transformation was completely successful.<br> Cultured them for subsequent experiments. </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 Aug 22</time> </li> </ul> <div class="step-recipe"> K592010, K592012, K864401, B0015 </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>Thu Aug 22</time> </li> </ul> <div class="step-recipe"> K592010, K592012, K864401<br> 100b up, 200b dn, KOD Plus NEO </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>Thu Aug 22</time> </li> </ul> <div class="step-recipe"> Cut B0015 with EcoRI, XbaI (using 10xM buffer)<br> Cut K592010, K592012, K864401 with EcoRI, SpeI (using 10xH buffer) </div> </div> </div> </li> | ||
| + | <li class="step"> <div class="step-tile"> <h2 class="step-title">Antarctic phosphatase</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"> 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>Thu Aug 22</time> </li> </ul> <div class="step-recipe"> Ligate B0015 with K592010, K592012, K864401 </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>Thu Aug 22</time> </li> </ul> <div class="step-recipe"> (K592010, K592012, K864401)-B0015<br> each of 5µL DNA to DH5α </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>Fri Aug 23</time> </li> </ul> <div class="step-recipe"> R0040-B0034 from our stock<br> 100b up, 200b dn, KOD Plus NEO </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>Fri Aug 23 and Sat Aug 24</time> </li> </ul> <div class="step-recipe"> (K592010, K592012, K864401)-B0015 </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>Sat Aug 24</time> </li> </ul> <div class="step-recipe"> (K592010, K592012, K864401)-B0015 </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 Aug 25</time> </li> </ul> <div class="step-recipe"> (K592010, K592012, K864401)-B0015 </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 K592010-B0015 with EcoRI, XbaI (using 10xM buffer)<br> Cut R0040-B0034 with EcoRI, SpeI (using 10xH 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"> (K592010, K592012, K864401)-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 25</time> </li> </ul> <div class="step-recipe"> Ligate (K592010, K592012, K864401)-B0015 with R0040-B0034 </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>Sun Aug 25</time> </li> </ul> <div class="step-recipe"> R0040-B0034-(K592010, K592012, K864401)-B0015<br> each of 5µL 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>Wed Sep 4</time> </li> </ul> <div class="step-recipe"> R0040-B0034-(K592010, K592012, K864401)-B0015 </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 Sep 5</time> </li> </ul> <div class="step-recipe"> R0040-B0034-(K592010, K592012, K864401)-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> | ||
Revision as of 05:39, 27 September 2013
Maestro E.coli
Notebook
-
Start
-
Get chromoprotein DNA
Great thanks to iGEM Uppsala! -
Transformation
K592010, K592012, K864401
each of 1µL DNA to DH5α -
Colony PCR & Culture
K592010, K592012, K864401, B0015
We confirmed transformation was completely successful.
Cultured them for subsequent experiments. -
Mini-prep
K592010, K592012, K864401, B0015 -
PCR & PCR purification
K592010, K592012, K864401
100b up, 200b dn, KOD Plus NEO -
Digestion
Cut B0015 with EcoRI, XbaI (using 10xM buffer)
Cut K592010, K592012, K864401 with EcoRI, SpeI (using 10xH buffer) -
Antarctic phosphatase
B0015 -
Ligation
Ligate B0015 with K592010, K592012, K864401 -
Transformation
(K592010, K592012, K864401)-B0015
each of 5µL DNA to DH5α -
PCR & PCR purification
R0040-B0034 from our stock
100b up, 200b dn, KOD Plus NEO -
Colony PCR
(K592010, K592012, K864401)-B0015 -
Culture
(K592010, K592012, K864401)-B0015 -
Mini-prep
(K592010, K592012, K864401)-B0015 -
Digestion
Cut K592010-B0015 with EcoRI, XbaI (using 10xM buffer)
Cut R0040-B0034 with EcoRI, SpeI (using 10xH buffer) -
Antarctic phophatase
(K592010, K592012, K864401)-B0015 -
Ligation
Ligate (K592010, K592012, K864401)-B0015 with R0040-B0034 -
Transformation
R0040-B0034-(K592010, K592012, K864401)-B0015
each of 5µL DNA to DH5α -
Culture
R0040-B0034-(K592010, K592012, K864401)-B0015 -
Mini-prep
R0040-B0034-(K592010, K592012, K864401)-B0015 -
Complete!
