User:DavidC/20 September 2009

From 2009.igem.org

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Contents

Sunday the 20th

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:

SU2009(1).png


Second: Quantization of P1003; C0012 and B0014:

SU2009(2).png



DNA purification

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

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