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Uppsala-Sweden/14 July 2011
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| + | Re-streak pSB1C3-ccaS_EcoRI, pGEM-amilGFP and pSB4A5. We were supposed to re-streak pSB1A3-amilCP and pSB1A3-PcpcG2 as well, but the transformants had no colonies at all. Suspect wrong plasmid backbone or no ligase added during the cloning attempt. To investigate this we did a transformation of the plasmid backbone (PSB1A3) and plated it on plates containing antibiotics ampicillin, chloramphenicol and kanamycin. We did plasmid prep of pSB1AK3-B1001, pSB1A2-R0011, pSB3T5-backbone, pSB4K5 backbone. New agar plates with antibiotics ampicillin, chloramphenicol and kanamycin, just made in the morning. Assembly of chromophores, step Cloning of tetR inverter into new backbone. Cloning of pSB1A3-amilCP and pSB1A3-PcpcG2 again. Overnight culture of pSB1A3 from the strain collection to test whether it’s right. | ||
| + | The sequencing result arrived today around lunch time! RBS-pcyA strains we sent for sequencing were good. ccaR clone number 2 is better than clone number 1. Clone 1 of RBS-ho1 will be used in further assembly steps. | ||
Latest revision as of 20:27, 20 July 2011
Welcome to Uppsala-SwedeniEM '2011
Notebook
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2011-07-14
Re-streak pSB1C3-ccaS_EcoRI, pGEM-amilGFP and pSB4A5. We were supposed to re-streak pSB1A3-amilCP and pSB1A3-PcpcG2 as well, but the transformants had no colonies at all. Suspect wrong plasmid backbone or no ligase added during the cloning attempt. To investigate this we did a transformation of the plasmid backbone (PSB1A3) and plated it on plates containing antibiotics ampicillin, chloramphenicol and kanamycin. We did plasmid prep of pSB1AK3-B1001, pSB1A2-R0011, pSB3T5-backbone, pSB4K5 backbone. New agar plates with antibiotics ampicillin, chloramphenicol and kanamycin, just made in the morning. Assembly of chromophores, step Cloning of tetR inverter into new backbone. Cloning of pSB1A3-amilCP and pSB1A3-PcpcG2 again. Overnight culture of pSB1A3 from the strain collection to test whether it’s right.
The sequencing result arrived today around lunch time! RBS-pcyA strains we sent for sequencing were good. ccaR clone number 2 is better than clone number 1. Clone 1 of RBS-ho1 will be used in further assembly steps.
