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Team:UNIPV-Pavia/Notebook/Week5Jun
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== <html><font class="dayw_style">June, 29th</font></html> == | == <html><font class="dayw_style">June, 29th</font></html> == | ||
| + | *We read that using primers VR/VF2 to PCR B0010 will result in excess bands, as documented by Samantha Burke in http://partsregistry.org/Problems_with_PCR_using_VR/VF2 . Maybe our extra bands were due to unwanted annealing between VR primer and B0015 which contains B0010. | ||
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| + | *In the future we will perform the screening of the ligations either through digestion or colony PCR taking carefully into account of their expected length and their potential non-specific primer binding sites. | ||
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| + | *We picked 4 colonies from A8 plate and 10 colonies from A9 plate (both stored at +4°C) and infected 1 ml of LB + Amp. We incubated the cultures at 37°C, 220 rpm for 5 and 1/2 hours. | ||
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| + | *Glycerol stocks for the grown cultures. | ||
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| + | *We re-filled the remaining 250 ul of bacterial culture with 5 ml of LB + Amp and incubated the cultures overnight at 37°C, 220 rpm. Tomorrow we will repeat the screening on these plates through miniprep/digestion! | ||
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== <html><font class="dayw_style">June, 30th</font></html> == | == <html><font class="dayw_style">June, 30th</font></html> == | ||
Revision as of 08:44, 29 June 2009
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Week from June 29th, to June 30th, 2009
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June, 29th
- We read that using primers VR/VF2 to PCR B0010 will result in excess bands, as documented by Samantha Burke in http://partsregistry.org/Problems_with_PCR_using_VR/VF2 . Maybe our extra bands were due to unwanted annealing between VR primer and B0015 which contains B0010.
- In the future we will perform the screening of the ligations either through digestion or colony PCR taking carefully into account of their expected length and their potential non-specific primer binding sites.
- We picked 4 colonies from A8 plate and 10 colonies from A9 plate (both stored at +4°C) and infected 1 ml of LB + Amp. We incubated the cultures at 37°C, 220 rpm for 5 and 1/2 hours.
- Glycerol stocks for the grown cultures.
- We re-filled the remaining 250 ul of bacterial culture with 5 ml of LB + Amp and incubated the cultures overnight at 37°C, 220 rpm. Tomorrow we will repeat the screening on these plates through miniprep/digestion!
June, 30th
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Previous Week
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Next Week
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