User:DavidC/12 October 2009
From 2009.igem.org
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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
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.