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JULY: WEEK 5
July, 25th
Red colonies were observed from E36 plates, suggesting that the transformation was successful. A colony was picked form the plate. Digestions of previously purified plasmids were performed for ligations:
Reactions were incubated at 37°C for three hours while a small-size and a medium-size agarose gel were prepared according to protocols. In the afternoon gel electrophoresis was performed. Immagine della corsa su gel After gel extraction, digested DNA was quantified:
Previously purified plasmids carrying either E17, or E18, or E19 or E20 part were inoculated in order to amplify them for the subsequent transformation in MGZ1 cells (E' GIUSTO?): July, 26th
Plasmids containing either E17, or E18, or E19, or E20 or E36 part were extracted with MiniPrep kit from ON saturated cultures; purified DNA was quantified with NanoDrop Spectrophotometer:
Digestion of E36 part carrying plasmid was performed for ligation:
Digestions of previously purified plasmids were performed for ligations:
Reactions were incubated at 37°C for three hours while a small-size agarose gel was prepared according to protocols. In the afternoon gel electrophoresis was performed. Immagine della corsa su gel After gel extraction, digested DNA was quantified:
Then ligations were performed in a final volume of 10 μl:
July, 27th
E37, E38, E39, E40, E41, E42, E17, E18, E19 and E20 were transformed in 100 μl of MGZ1 competent cells according to protocols. Moreover a plasmid containing BBa_B0032 was transformed in order to test the transformation efficiency. Plates were incubated ON at 37°C. July, 28th
Colonies grew in every plate. Infact both 17-2, E18-2, E19-2 and E20-2 parts, whose DNA was recovered after MiniPrep extraction kit, both E37, E38, E39 and E40 parts showed grown colonies. We picked two colonies from all parts. As regards to E41 and E42 parts, we resolved to let them still grow.
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Team:UNIPV-Pavia/Calendar/July/settimana5
From 2011.igem.org
