Team:Paris/18 August 2009
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Cha.olivier (Talk | contribs) (→Lab work) |
Cha.olivier (Talk | contribs) (→Lab work) |
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The ligation that I have set up yesterday (pSB1A3 [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#P16 pSB1A3] w/ D13 [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#D13 D13] or D14 [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#D14 D14] ) didn't work so I decided to redigest PCR products (A11 (RBS-ompA signal) and A13 (TolRII). | The ligation that I have set up yesterday (pSB1A3 [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#P16 pSB1A3] w/ D13 [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#D13 D13] or D14 [https://2009.igem.org/Team:Paris/Freezer_Digestion_Products#D14 D14] ) didn't work so I decided to redigest PCR products (A11 (RBS-ompA signal) and A13 (TolRII). | ||
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''Mix : total volume = 30 uL'' | ''Mix : total volume = 30 uL'' |
Revision as of 09:40, 18 August 2009
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Brain work
edit please ^^
Lab work
PCR on colony w/ Soufifi
The ligation pLAC/RBS-TetR worked --> no clone on the negatif control and some clone on the "good" plates.
PCR on colony is required to check the validity of our clones.
Mix : total volume = 25uL
Super Mix 2X: 12,5 uL VF2 (10uM): 0,5 uL VR (10 uM): 0,5 uL H20 : 6,5 uL diluted bacteria : 5 uL
Tm = 55°C Elongation time = 1 min
Digestion
The ligation that I have set up yesterday (pSB1A3 pSB1A3 w/ D13 D13 or D14 D14 ) didn't work so I decided to redigest PCR products (A11 (RBS-ompA signal) and A13 (TolRII).
Mix : total volume = 30 uL
For A11:
DNA = 15 uL
10X buffer: 3 uL
Enz1: 1 uL
Enz2: 1 uL
BSA: 0,5 uL
H2O: 9,5 uL
For A11:
DNA = 10 uL 10X buffer: 3 uL
Enz1: 1 uL
Enz2: 1 uL
BSA: 0,5 uL
H2O: 14,5 uL
To do list
edit please ^^