User:DavidC/28 September 2009

DNA extraction
DNA extraction with miniprep (promega).

Samples:

BBa_P1003 + BBa_B0014; BBa_C0012 + BBa_B0014; BBa_B0014 + BBa_J23100 + BBa_B0030; BBa_R0010 + BBa_B0014. BBa_I712669 + BBa_B0014

Final volume = 30µL

Restriction digest
Restriction digest of I712669+B0014 by EcoRI and SpeI (4806bp):

DNA (miniprep) = 30µL Buffer 2 (NEB) =5µL H20 = 13µL Eco RI (NEB) = 1µL Spe I (NEB) = 1µL 1 hour of incubation at 37°C.

Restriction digest of P1003 by EcoRI and XbaI:

DNA (miniprep) = 30µL Buffer 2 (NEB) =5µL H20 = 13µL Eco RI (NEB) = 1µL Xba I (NEB) = 1µL 1 hour of incubation at 37°C.

Restriction digest of C0012 + B0014 by XbaI and PstI:

DNA (miniprep) = 30µL Buffer 2 (NEB) =5µL H20 = 13µL Xba 1 (NEB) = 1µL Pst 1 (NEB) = 1µL 1 hour of incubation at 37°C.

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

Samples: BBa_P1003, BBa_I712669 + BBa_B0014, BBa_C0012 + BBa_B0014.



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