IGEM:Peking/2007/Count-Conjugation-Notebook/2007-7-21

Ligation e0040->pSB3k3

 * use T4 Ligase to ligate.
 * Ligation system contains (totally 20uL): 4uL pSB3K3(I7100), 3uLEGFP(E0040),1uL 10XT4L buffer, 1uL T4 L, 1uL dH20.

PCR crRNA

 * PCR system contains (totally 10uL): 0.5uL P1, 0.5uL P2, 2.5uL 10*b, dNTP 2uL, Taq 0.25uL, dH20 19.25uL.
 * PCR condition:
 * 1) 94℃ 5min
 * 2) 94℃ 30s
 * 3) 60℃ 30s
 * 4) 72℃ 30s
 * 5) Go to step 2 for 29 times
 * 6) 72℃ 10min
 * 7) 4℃  end