Team:Brown-Stanford/Lab/Protocol/BiobrickRestrictionDigest

Biobrick Restriction Digest

(Taken from [http://openwetware.org/wiki/Restriction_digest openwetware.org]; the protocol was subsequently edited for restriction of [http://partsregistry.org/Part:BBa_K656013 Part BBa_K656013] and [http://partsregistry.org/Part:pSB1C3 pSB1C3])

Reagents for BBa_K656013 digestion

• 4.6 µL nuclease-free water
• 27 µL purified PCR product
• 4 µL appropriate NEBuffer
• .4 µL BSA
• 2 µL each restriction enzyme

Treatment for BBa_K656013

1. Combine all reagents in a single PCR tube
2. Incubate at 37˚C for 3 hours

Reagents for pSCB1C3 digestion

• 27.4 µL nuclease-free water
• 3.6 µL miniprepped DNA sample
• 4 µL appropriate NEBuffer
• .4 µL BSA
• 1 µL each restriction enzyme

Treatment for pSCB1C3"

1. Combine all reagents in a single PCR tube
2. Incubate at 37˚ for 1 hour