Ligation of:

1. E7(I)-Terminator(V)
2. LsrA(V)-Lysis(I)
3. Lysis(V)-Terminator(I)

• 900-930:Hung: digestion of E7-I(E/S), Terminator-V(E/X), GFP (E/S) to compare the bands with E7, LsrA-V(S/P), Lysis-I (X/P), Lysis-V(S/P), Terminator-I(X/P). Incubation for 2.5 hours.
• 1200-1230: Lu Chao: Minelute PCR purification.
• 1230-1430: Lu Chao: gel loading+running+staining.
• 1430-1600: gel extraction + nanodrops. All the bands needed were shown under UV!
• 1600-1645: ligation.
• 1645-1715: MinElute for ligation mixtures.
• 1715-1900: transformation into Top10.