User:DavidC/20 September 2009

Restriction digest
P1003 (= 2,5µg/µL) restriction digest by EcoRI and SpeI:

DNA (10µg final) = 4µL Buffer H (TAKARA) =2µL H20 = 12µL EcoR1 (TAKARA) = 1µL Spe 1 (TAKARA) = 1µL > Incubation 1h at 37°C.

Calf intestine phosphatase (CIP) processing: Restriction digest mix = 20µL CIP Buffer (TAKARA) = 5µL H20 = 24µL CIP (TAKARA) = 1µL 1h at 37°C

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

First: Purification of P1003 fragment:



Second: Quantization of P1003; C0012 and B0014:



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