Line 210: | Line 210: | ||
<div id="4" class="yolo" style="display:none;"> | <div id="4" class="yolo" style="display:none;"> | ||
<p> | <p> | ||
− | <br> | + | <br>• PCR: 9, 10, 16, 29, 30, 31, 32, 33, 34, 35, 36, 41, 42, 43</br> |
Each PCR was loaded on a preparative gel and cut out before undergoing PCR-purification. | Each PCR was loaded on a preparative gel and cut out before undergoing PCR-purification. | ||
− | <br><br> | + | <br><br>• Golden Gate Assembly: BB1_16, BB1_17, BB1_18, BB1_20, BB1_21, BB1_22, BB1_23, BB1_24, BB1_25, BB2_17, BB2_23, BB2_24, BB3_01 (unsuccessful), BB3_02, , BB3_04, BB3_05, BB3_06 (doubtful), BB3_08</br> |
GG-products are then transformed into competent cells. Colonies with specific antibiotic resistances were selected out by using antibiotic-containing media. The next day, positives clones were confirmed via a OneTaq colony PCR and a gel electrophoresis before a Miniprep was made.</br> | GG-products are then transformed into competent cells. Colonies with specific antibiotic resistances were selected out by using antibiotic-containing media. The next day, positives clones were confirmed via a OneTaq colony PCR and a gel electrophoresis before a Miniprep was made.</br> | ||
− | <br> | + | <br>• Yeast S266C made electro-competent</br> |
− | <br> | + | <br>• Sent for Sequencing: BB1_16, BB1_17, BB2_18, BB3_02, BB3_05, CRISPR-PPP</br> |
</p></div> | </p></div> | ||
Line 222: | Line 222: | ||
<div id="5" class="yolo" style="display:none;"> | <div id="5" class="yolo" style="display:none;"> | ||
<p> | <p> | ||
− | <br>•</br> | + | <br>• PCR: 41, 42, 45, 52, 53, 56, 57</br> |
− | <br>•</br> | + | Each PCR was loaded on a preparative gel and cut out before undergoing PCR-purification. |
− | <br>•</br> | + | <br><br>• Golden Gate Assembly: BB1_19, BB2_19, BB2_25, BB2_26, BB2_27, BB2_28, BB2_29, BB2_30, BB2_31, BB2_32, BB2_33, BB2_34, BB3_01(unsuccessful), BB3_02, BB3_03(unsuccessful), BB3_06, BB3_10, BB3_13-15, BB3_24</br> |
− | <br>•</br> | + | GG-products are then transformed into competent cells. Colonies with specific antibiotic resistances were selected out by using antibiotic-containing media. The next day, positives clones were confirmed via a OneTaq colony PCR and a gel electrophoresis before a Miniprep was made.</br> |
− | <br>•</br> | + | <br>• Preculture of DH10B <i>E. coli</i> strain made chemically competent</br> |
+ | <br>• Yeast cells transformed with the Cas9 plasmid</br> | ||
+ | <br>• Sent for Sequencing: BB1_19, CRISPR 2, CRISPR 3, BB3_12, BB3_13, BB3_14, BB2_34</br> | ||
</p></div> | </p></div> | ||
Revision as of 20:01, 28 October 2017
Notebook.
All protocols we used for the methods can be found here:
Protocol
Week 1 (03/07-07/07)
Week 2 (10/07-14/07)
Week 3 (17/07-21/07)
Week 4 (24/07-28/07)
Week 5 (31/07-04/08)
Week 6 (07/08-11/08)
Week 7 (14/08-18/08)
Week 8 (21/08-25/08)
Week 9 (28/08-01/09)
Week 10 (04/09-08/09)
Week 11 (11/09-15/09)
Week 12 (18/09-22/09)
Week 13 (25/09-29/09)