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Team:Brown-Stanford/Lab/Protocol/BiobrickRestrictionDigest
From 2011.igem.org
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(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]) | (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''' | ||
| + | # Combine all reagents in a single PCR tube | ||
| + | # 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" | ||
| + | # Combine all reagents in a single PCR tube | ||
| + | #Incubate at 37˚ for 1 hour | ||
Revision as of 08:54, 28 September 2011
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
- Combine all reagents in a single PCR tube
- 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"
- Combine all reagents in a single PCR tube
- Incubate at 37˚ for 1 hour
