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<br>• PCR: 27, 28</br> | <br>• PCR: 27, 28</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>• Golden Gate Assembly: BB1_14, BB1_15, BB2_01 (unsuccessful, even with new BBa_23101 promoter), BB2_05 (unsuccessful), BB2_06, BB2_07, BB2_08, BB2_09, BB2_10, BB2_12, BB2_14, BB2_15, BB2_16, | + | <br><br>• Golden Gate Assembly: BB1_14, BB1_15, BB2_01 (unsuccessful, even with new BBa_23101 promoter), BB2_05 (unsuccessful), BB2_06, BB2_07, BB2_08, BB2_09, BB2_10, BB2_12, BB2_14, BB2_15, BB2_16, C-PPP_01 (unsuccessful)</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>• Sent for Sequencing: BB1_14, BB1_15, BB2_01, BB2_05, BB2_06, BB2_07, BB2_08, BB2_09, BB2_10, BB2_12</br> | <br>• Sent for Sequencing: BB1_14, BB1_15, BB2_01, BB2_05, BB2_06, BB2_07, BB2_08, BB2_09, BB2_10, BB2_12</br> | ||
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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>• Yeast S266C made electro-competent</br> | <br>• Yeast S266C made electro-competent</br> | ||
− | <br>• Sent for Sequencing: BB1_16, BB1_17, BB2_18, BB3_02, BB3_05, | + | <br>• Sent for Sequencing: BB1_16, BB1_17, BB2_18, BB3_02, BB3_05, C-PPP_01</br> |
</p></div> | </p></div> | ||
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<br>• Preculture of DH10B <i>E. coli</i> strain made chemically competent</br> | <br>• Preculture of DH10B <i>E. coli</i> strain made chemically competent</br> | ||
<br>• Yeast cells transformed with the Cas9 plasmid</br> | <br>• Yeast cells transformed with the Cas9 plasmid</br> | ||
− | <br>• Sent for Sequencing: BB1_19, | + | <br>• Sent for Sequencing: BB1_19, BB2_34, BB3_12, BB3_13, BB3_14, C-PPP_02, C-PPP_03</br> |
</p></div> | </p></div> | ||
Revision as of 20:20, 28 October 2017
Notebook.
All protocols we used for the methods can be found here:
Protocol
For further information regarding our Golden Gate Assembly products, please refer to our part collection site.
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)