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Team:UNIPV-Pavia/Notebook/Week3Aug
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== <html><font class="dayw_style">August, 21st</font></html> == | == <html><font class="dayw_style">August, 21st</font></html> == | ||
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| + | *Colony PCR for A14 (5 colonies) and A16 (5 colonies) plates. Colonies were inoculated in 1 ml of LB + Amp and let grow waiting for the end of the reaction. | ||
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| + | *Electrophoresis for PCR results. | ||
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| + | *Gel results: | ||
| + | **A14 - 3rd colony seems the most pure and it had the expected length for ligated plasmid. | ||
| + | **A16 - reaction worked only on A16-1, but it was negative. | ||
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| + | *We decided to keep A14-3 (positive at PCR) and A16-4 (randomly chosen) for digestion screening: we prepared a glycerol stock for them and re-filled the remaining 250 ul of bacteria with 4 ml of LB + Amp. | ||
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| + | *We incubated these cultures at 37°C, 220 rpm overnight. | ||
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| + | *We picked 2 colonies from B7, 5 colonies from B8 and 1 colony from pSB3K3 plates. We inoculated them in 1 ml of LB + Kan and incubated them at 37°C, 220 rpm for 5 hours and 1/2. Then we prepared glycerol stocks and re-filled the remaining 250 ul with 4 ml of LB + Kan. | ||
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| + | *NOTE: we decided not to perform PCR on B7 and B8 plates because of the large size of positive inserts. | ||
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Revision as of 11:05, 23 August 2009
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Week from August 17th, to August 23rd, 2009
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August, 17th
August, 18th
August, 19th
August, 20th
August, 21st
- Colony PCR for A14 (5 colonies) and A16 (5 colonies) plates. Colonies were inoculated in 1 ml of LB + Amp and let grow waiting for the end of the reaction.
- Electrophoresis for PCR results.
- Gel results:
- A14 - 3rd colony seems the most pure and it had the expected length for ligated plasmid.
- A16 - reaction worked only on A16-1, but it was negative.
- We decided to keep A14-3 (positive at PCR) and A16-4 (randomly chosen) for digestion screening: we prepared a glycerol stock for them and re-filled the remaining 250 ul of bacteria with 4 ml of LB + Amp.
- We incubated these cultures at 37°C, 220 rpm overnight.
- We picked 2 colonies from B7, 5 colonies from B8 and 1 colony from pSB3K3 plates. We inoculated them in 1 ml of LB + Kan and incubated them at 37°C, 220 rpm for 5 hours and 1/2. Then we prepared glycerol stocks and re-filled the remaining 250 ul with 4 ml of LB + Kan.
- NOTE: we decided not to perform PCR on B7 and B8 plates because of the large size of positive inserts.
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Previous Week
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Next Week
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