Team:UNIPV-Pavia/Calendar/July/settimana1
From 2010.igem.org
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{| border='1' align='center' | {| border='1' align='center' | ||
- | || '''name''' || '''description''' || '''growth conditions''' | + | || '''Culture name''' || '''description''' || '''growth conditions''' |
|- | |- | ||
|| BT340 || pCP20 plasmid || LB+Amp LC 30°C,LB+Amp HC 30°C | || BT340 || pCP20 plasmid || LB+Amp LC 30°C,LB+Amp HC 30°C | ||
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Cultures were MiniPrepped with the following quantifications (NanoDrop): | Cultures were MiniPrepped with the following quantifications (NanoDrop): | ||
{| border='1' align='center' | {| border='1' align='center' | ||
- | || ''' | + | || '''Culture name''' ||'''quantification''' |
|- | |- | ||
|| I7-1 || 83,3 ng/ul | || I7-1 || 83,3 ng/ul | ||
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|| PhaP-2 || 13,6 ng/ul | || PhaP-2 || 13,6 ng/ul | ||
|} | |} | ||
+ | |||
+ | All cultures, except PhaP-1 and PhaP-2 were digested. Digestion of: | ||
+ | |||
+ | {| border="1" align='center' | ||
+ | | ''Culture'' || ''Kind'' || ''Final reaction volume (ul) '' || ''DNA (ul)'' || ''H20 (ul)'' || ''Enzyme 1'' || ''Enzyme 2'' || ''Buffer H'' | ||
+ | |- | ||
+ | | I7-1 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |- | ||
+ | | I7-2 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |- | ||
+ | | I8-1 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |- | ||
+ | | I8-2 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |- | ||
+ | | I9-1 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |- | ||
+ | | I9-2 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |- | ||
+ | | I10-1 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |- | ||
+ | | I10-2 || Screening || 25 || 2 || 19,5 || 0,5 EcoRI || 0,5 PstI || 2,5 | ||
+ | |} | ||
+ | |||
+ | Digestion was performed at 37°C for 3 hours. Digestions were gel run. For I9 and I10 clones all colonie were correct, while for I7 and I8 extra bands could be observed. Since preliminar results of TECAN test didn't provide encouraging results, we decided to select other 3 colonies both for I7 and I8 and to repeat the screening, while I9 and I10 were correct at this screening, so we choosed I9-1 and I10-1 as definitive I9 and I10 parts. I9 and I10 will be further screened next week. | ||
+ | |||
+ | |||
+ | ________________________ | ||
+ | TECAN TEST | ||
+ | ________________________ | ||
[[Image:Unipv_screening_I7_I8_I9_I10.jpg|300px|thumb|center|Screening of I7, I8, I9, I10]] | [[Image:Unipv_screening_I7_I8_I9_I10.jpg|300px|thumb|center|Screening of I7, I8, I9, I10]] | ||
+ | |||
+ | |||
+ | |||
All strains received were on blotting paper disks, that were placed on an LB agar plates (with/without antiobiotic, see growth conditions), resuspended wih 80ul LB and then streaked. After plate streaking, disks were trasnferred in falcon containing liquid LB. | All strains received were on blotting paper disks, that were placed on an LB agar plates (with/without antiobiotic, see growth conditions), resuspended wih 80ul LB and then streaked. After plate streaking, disks were trasnferred in falcon containing liquid LB. |
Revision as of 12:02, 5 July 2010
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