IGEM:Peking/2007/Switch-Notebook/2007-8-14

Precipitate for retraction
SulA, Rec, SD1-3, SS1-2, GFP-pLX007 (^^ with XbaI & XhoI)

Ligate with GFP-pLX007
SulA, Rec, SD1-3, SS1-2

Colony PCR test of Prec, SD2-3, SS1-3-(SDY-EGFP-pLX007)
Number of colonies grew up: No. -- Prec   0 SD2    0 SD3    1 SS1  >50 SS2    2 SS3   14

Only SS1 is right. 4 colonies are select for incubation.

Colony PCR test of T1T2-(Tet F-lacZ-pLX010)
22 colonies are selected for incubation.

PCR of RD1
Retract through electropheresis.

Double digestion of RD1
10uL  10*H buffer 2uL  SpeI 2uL  XhoI 15uL  RD1 71uL  ddH2O --- 100uL  total