Paris/September 13
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(→DapA Wanner deletion: pKD3/pKD4-DapA homologies PCR) |
(→DapA Wanner deletion: pKD3/pKD4-DapA homologies PCR) |
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We diluted 1/100 overnight culture at 30°C of MG1655 transformed by pKD46: 50µL into 5mL LB ampicilline.<br> | We diluted 1/100 overnight culture at 30°C of MG1655 transformed by pKD46: 50µL into 5mL LB ampicilline.<br> | ||
At OD = 0.1, we induced pKD46 expression by addition of arabinose (10mM final). <br> | At OD = 0.1, we induced pKD46 expression by addition of arabinose (10mM final). <br> | ||
| - | At OD = 0.4, we prepared cells to be electrocompetent.<br> | + | At OD = 0.4, we prepared cells to be electrocompetent. (see [[Paris/PROTOCOLS#Recombineering.2FLambda_red-mediated_gene_replacement|Protocol]])<br> |
Then we transformed the bacteria with PCR product from yesterday (see [[Paris/September_12|September 12]]). | Then we transformed the bacteria with PCR product from yesterday (see [[Paris/September_12|September 12]]). | ||
Latest revision as of 14:50, 14 September 2007
DapA Wanner deletion: pKD3/pKD4-DapA homologies PCR
We diluted 1/100 overnight culture at 30°C of MG1655 transformed by pKD46: 50µL into 5mL LB ampicilline.
At OD = 0.1, we induced pKD46 expression by addition of arabinose (10mM final).
At OD = 0.4, we prepared cells to be electrocompetent. (see Protocol)
Then we transformed the bacteria with PCR product from yesterday (see September 12).
Sequencing
The following plasmids were sent for sequencing rxn:
(name of the plasmid+oligo)
DB_L31.1+O18 DB_L31.1+O19 DB_L31.3+O18 DB_L31.3+O19 DB_L32.1+O18 DB_L32.1+O19 DB_L32.2+O18 DB_L32.2+O19 DB_L34.1+O18 DB_L34.1+O19 DB_L34.2+O18 DB_L34.2+O19 DB_L35.1+O18 DB_L35.1+O19 DB_L35.2+O18 DB_L35.2+O19 DB_L36.1+O18 DB_L36.1+O19 DB_L36.2+O18 DB_L36.2+O19 DB_L37.1+O18 DB_L37.1+O19 DB_L37.1+O26 DB_L37.2+O18 DB_L37.2+O19 DB_L37.2+O26 DB_L39.1+O18 DB_L39.1+O19 DB_L39.1+O26 DB_L39.2+O18 DB_L39.2+O19 DB_L39.2+O26 DB_L40.1+O18 DB_L40.1+O19 DB_L40.1+O25 DB_L40.2+O18 DB_L40.2+O19 DB_L40.2+O25 DB_L41.1+O18 DB_L41.1+O19 DB_L41.1+O24 DB_L42.1+O18 DB_L42.1+O19 DB_L42.1+O24 DB_L42.1+O22 DB_L42.1+O23 DB_L42.2+O18 DB_L42.2+O19 DB_L42.2+O24 DB_L42.2+O22 DB_L42.2+O23 DB_L42.3+O18 DB_L42.3+O19 DB_L42.3+O24 DB_L42.3+O22 DB_L42.3+O23 DB_L43.1+O18 DB_L43.1+O19 DB_L43.1+O25 DB_L43.2+O18 DB_L43.2+O19 DB_L43.2+O25 DB_L43.3+O18 DB_L43.3+O19 DB_L43.3+O25 DB_MPT5.1+O41 DB_MPT5.2+O41
