User:DavidC/29 September 2009

Ligation between D protein and adenovirus 5 penton base
Restriction digest of D protein by Spe I (380bp): DNA (PCR with bio-brick primers) = 5µL Buffer M (TAKARA) =2µL BSA (TAKARA) = 2µL <br H20 = 10µL Spe 1 (TAKARA) = 1µL 1 hour of incubation at 37°C.

Restriction digest of adenovirus 5 penton base by XbaI (1715bp):

DNA (PCR with bio-brick primers) = 5µL Buffer M (TAKARA) =2µL BSA (TAKARA) = 2µL <br H20 = 10µL Xba 1 (TAKARA) = 1µL 1 hour of incubation at 37°C.

DNA electrophoresis
85 Volt, 15 minutes. 105 Volt, 40 minutes. Ladder fermentas 1 Kb.

Samples: D protein cut by SpeI (D prt), ADV5pb protein cut by XbaI.



DNA purification
Kit Qiagen “gel extraction kit”, final volume = 50µL.