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INSA-Lyon/29 September 2010
From 2010.igem.org
September 29th
DNA extraction of Curli, 8C/14K, 24A, 14K, 18A, L2/1N, phasin, phasin-intein
Digestion for a standard ligation of 18A, 24A and phasin
Agarose gel electrophoresis for weight verification :
- 8C/14K : OK
- curli : nothing, to do again
- 24A : OK
- 14K : not enough DNA, to do again
- 18A : OK
- L2/1N : nothing, to do again
- phasin : nothing, to do again
- phasin-intein : OK
Culture on plate of 14K, L2/1N, phasin, I0500, 4O, 10L, curli, 8C 24A and 10 clones from the ligation 1F/24A+14K of yesterday
Liquid culture of I0500, 4O and 10L
Collection for Curli M.Gene and 8C/14K: 750µl of each culture in glycerol (750µl). Stored at -80°C.
PCR PSB program on pSB1T3 and pSB1C3 PCR XT gel (2010/09/27) with five times more dNTP (no more Roche dNTP), PCR products are stored at -20°C.
Transformation in XL-Blue competent cells (Stratagene protocol): 70µl ligation ompR-pSB1C3 in 200µl cells; 20µl 1F/24/14K in 100µl cells; 20µl sterile water in 100µl cells.
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