User:DavidC/12 October 2009

Monday the 12th
DNA extraction:

Extraction of p53 from E.coli DH5alpha using a maxiprep (macherey nagel) (2 times).

Minipreps: BBa_B0030; BBa_C0012; BBa_B0014; BBa_ B0030 + BBa_C0012 + BBa_B0014.

Restriction digest of BBa_B0030 + BBa_C0012 + BBa_B0014 by XbaI and PstI
DNA (miniprep) = 30µL Buffer H (TAKARA) = 4µL H20 = 4µL PstI (TAKARA) = 1µL XbaI (TAKARA) = 1µL 1 hour of incubation at 37°C.

Restriction digest of BBa_B0014 by EcoRI and XbaI
DNA (miniprep) = 30µL Buffer H (TAKARA) = 4µL H20 = 4µL PstI (TAKARA) = 1µL XbaI (TAKARA) = 1µL 1 hour of incubation at 37°C.

Restriction digest of BBa_C0012 by EciRI and XbaI
DNA (miniprep) = 30µL Buffer H (TAKARA) = 4µL H20 = 4µL PstI (TAKARA) = 1µL XbaI (TAKARA) = 1µL 1 hour of incubation at 37°C.

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

Samples:

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