Team:Paris/12 August 2009

NoteBook
August 12th  

Lab work
Ligation D15 (2xterminator E/S) and D16 (pLac E/X) with vector psb2k3

DO : D15=1,19µg/mL  D16=1,75µg/mL

1h Home temperature

Transformation classic protocol with DH5a

Purification
 * Purification of A10(ClyA) (PCR at 55° during 30sec) (using 1% agarose during 25 min)
 * A10=A10 987pb
 * A10 purificated
 * Purification of A10 work

PCR Gel verification 1% aga gel after PCR under different conditions for A18 (OmpA-Linker from Varsaw). Expected weight : 525bp. Obviously, something has gone wrong. Next step will be different temperature gradients testing for PCR and/or new matrix.

Transformation Classic protocol with DH5a, and pSB2K3 from iGEM DNA plates (1/7C). ON culture on LB + Kan.

To be checked tomorrow !

ON culture for Minipreps tomorrow pSB1A3, pLAC, RBSTetR, OmpA-L, Three ligation clones, ON culture in LB + Abiotics. 37°C with stiring.

ON PCR A20 : 49, 51 and 53°C PCR with the plasmid for OmpA-L. Overnigth. 15s elongation time.

All Manip Comming soon

 