Team:Brown-Stanford/Lab/Protocol/BiobrickRestrictionDigest

From 2011.igem.org

Brown-Stanford
iGEM

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