Team:Aachen/Notebook/Wetlab/June
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__NOTOC__ | __NOTOC__ | ||
+ | {{CSS/Main}} | ||
+ | {{Team:Aachen/Stylesheet}} | ||
{{Team:Aachen/Header}} | {{Team:Aachen/Header}} | ||
+ | |||
+ | <html> | ||
+ | <center> | ||
+ | <ul class="menusmall-grid"> | ||
+ | |||
+ | <!-- <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/March" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to March</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/7/7a/Aachen_14-10-10_March_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> --> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/April" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to April</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/2/2d/Aachen_14-10-10_April_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/May" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to May</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/67/Aachen_14-10-10_May_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/June" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to June</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/1/1d/Aachen_14-10-10_June_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/July" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to July</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/1/19/Aachen_14-10-10_July_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/August" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to August</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/4/4e/Aachen_14-10-10_August_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/September" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to September</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/d/d4/Aachen_14-10-10_September_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/October" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to October</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/60/Aachen_14-10-10_October_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | </ul> | ||
+ | </center> | ||
+ | </html> | ||
= June = | = June = | ||
== 3rd == | == 3rd == | ||
- | * | + | * 34 BioBricks were transformed |
== 4th == | == 4th == | ||
- | * | + | * Preparation of consumables: |
- | ** fresh 50 | + | ** fresh 50% glycerol |
** new LB plates with cam and with kanamycin (kan) | ** new LB plates with cam and with kanamycin (kan) | ||
** 60 glas tubes | ** 60 glas tubes | ||
Line 15: | Line 89: | ||
** 100 mL steril glas beads for plating | ** 100 mL steril glas beads for plating | ||
- | * | + | * Master plates (6 clones per BioBrick) were made |
== 5th == | == 5th == | ||
- | * | + | * Colony PCRs on all transformed BioBricks were conducted |
*: → 2 clones from each master plate were picked | *: → 2 clones from each master plate were picked | ||
- | * | + | * Overnight cultures in freshly prepared 5 mL LB + cam were inoculated |
*: → 1 culture per BioBrick | *: → 1 culture per BioBrick | ||
== 6th == | == 6th == | ||
- | * | + | * 2 glycerol stocks of each BioBrick were made |
== 14th == | == 14th == | ||
'''PCR of E0030 to K1319000''' | '''PCR of E0030 to K1319000''' | ||
- | * Q5 and Phusion polymerase | + | * Q5 and Phusion polymerase were used |
<center> | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
- | ! | + | ! Parameter !! Duration !! Temp [°C] |
|- | |- | ||
- | | | + | | Denature||5:00||98 |
|- | |- | ||
- | | ''' | + | | '''Denature'''||00:30||98 |
|- | |- | ||
- | | ''' | + | | '''Anneal'''||00:30||55 |
|- | |- | ||
- | | ''' | + | | '''Elongate'''||00:50||72 |
|- | |- | ||
- | | | + | | Elongate||05:00||72 |
|} | |} | ||
</center> | </center> | ||
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== 17th == | == 17th == | ||
- | * | + | * Colony PCR on following constructs: |
<center> | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
|- | |- | ||
- | ! ID !! | + | ! ID !! Colony !! Product Length |
|- | |- | ||
| 1, 2 || K1319000 || 1041 | | 1, 2 || K1319000 || 1041 | ||
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|} | |} | ||
</center> | </center> | ||
- | → | + | → Anneling: 50°C |
- | → | + | → Elongation: 02:57 |
== 20th == | == 20th == | ||
- | * | + | * Colony PCR of K325108, K325218, C0062 |
<center> | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
- | ! | + | ! Parameter !! Duration !! Temp [°C] |
|- | |- | ||
- | | | + | | Denature||5:00||95 |
|- | |- | ||
- | | ''' | + | | '''Denature'''||00:30||95 |
|- | |- | ||
- | | ''' | + | | '''Anneal'''||00:30||49 |
|- | |- | ||
- | | ''' | + | | '''Elongate'''||03:15||72 |
|- | |- | ||
- | | | + | | Elongate||05:00||72 |
|} | |} | ||
</center> | </center> | ||
→ 30 cycles | → 30 cycles | ||
- | * PAGE: 30 min on 1.2 | + | * PAGE: 30 min on 1.2% agarose gel at 110 V |
→ weak bands for K325108 a & b at 4.5 kb and 9 kb respectively | → weak bands for K325108 a & b at 4.5 kb and 9 kb respectively | ||
== 23rd == | == 23rd == | ||
- | * | + | * Yesterdays colony PCR was repeated |
- | * | + | * Precultures for plasmid preps were inoculated |
** K1319000 → sequencing | ** K1319000 → sequencing | ||
** K592100 → BFP | ** K592100 → BFP | ||
Line 104: | Line 178: | ||
== 24th == | == 24th == | ||
* 8 plasmid preps were conducted | * 8 plasmid preps were conducted | ||
- | * | + | * Overnight cultures of K516132 and J23101.E0240 were inoculated |
== 25th == | == 25th == | ||
- | * | + | * Preculture for competent NEB10β cells was set up |
- | * | + | * Samples for sequencing were submitted |
- | * | + | * Master plates of transformed BioBricks were made |
- | * | + | * Overnight expression cultures of J23101.E0240 and K516132 were centrifuged and frozen |
- | * | + | * Fresh 50% glycerol was prepared |
== 26th == | == 26th == | ||
Line 119: | Line 193: | ||
== 27th == | == 27th == | ||
- | * | + | * Plasmid preps |
<center> | <center> | ||
{| class="wikitable" | {| class="wikitable" | ||
Line 142: | Line 216: | ||
|} | |} | ||
</center> | </center> | ||
- | * | + | * Two cryos stocks of each were prepared |
== 28th == | == 28th == | ||
- | * | + | * Sequencing |
- | * | + | * Transformation of 17 BioBricks |
== 30th == | == 30th == | ||
- | * | + | * Master plates |
+ | <html> | ||
+ | <center> | ||
+ | <ul class="menusmall-grid"> | ||
+ | |||
+ | <!-- <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/March" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to March</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/7/7a/Aachen_14-10-10_March_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> --> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/April" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to April</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/2/2d/Aachen_14-10-10_April_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/May" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to May</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/67/Aachen_14-10-10_May_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/June" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to June</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/1/1d/Aachen_14-10-10_June_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/July" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to July</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/1/19/Aachen_14-10-10_July_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/August" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to August</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/4/4e/Aachen_14-10-10_August_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/September" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to September</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/d/d4/Aachen_14-10-10_September_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | <li style="width:106px;margin-left: 12px;margin-right: 12px;" > | ||
+ | <a class="menulink" href="https://2014.igem.org/Team:Aachen/Notebook/Wetlab/October" style="color:black"> | ||
+ | <div class="menusmall-item menusmall-info" style="height:100px; width: 100px;" ><div class="menukachel" style="top: 30%; font-size: 16px;">Go to October</div></div> | ||
+ | <div class="menusmall-item menusmall-img" style="background: url(https://static.igem.org/mediawiki/2014/6/60/Aachen_14-10-10_October_iFG.png); norepeat scroll 0% 0% transparent; background-size:100%; height:100px; width: 100px;"> | ||
+ | </div> | ||
+ | </a> | ||
+ | </li> | ||
+ | |||
+ | </ul> | ||
+ | </center> | ||
+ | </html> | ||
{{Team:Aachen/Footer}} | {{Team:Aachen/Footer}} |
Latest revision as of 16:38, 17 October 2014
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