User:DavidC/29 September 2009
From 2009.igem.org
Contents |
Tuesday the 29th
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.