All Enzymes used for Bird of Prey
Below are the ten enzymes required to fully unlock the power of Bird of Prey and your CloneCards. For a more detailed tutorial, please check the handbook or overview chapter. While we recommend purchasing all enzymes at first, some will likely be used more than others depending on what you want to achieve. All of these enzymes are available at New England Biolabs.
AgeI
3' Flank of P Domain
Used to grab a whole Transcription Unit without the TUL domain or in a P domain modification event. We recommend the High Fidelity version of this enzyme.
AseI
The "Linearizer"
Sometimes your Bird of Prey Transcription Units will be as large or larger than the backbone itself. Make it easy with AseI and linearize your construct for easier building.
KasI
5' Flank of P | 3' Flank of EL
Used to move an entire Effector domain - either Mono-Effector or a Multi-Effector. We recommend the High Fidelity version of this enzyme.
NotI
3' Flank of the TUL Domain
Used to release a Mono-TU or a Multi-TU from the backbone, or building a multigene (XhoI/NotI). We recommend the High Fidelity version of this enzyme.
PvuI
5' Flank of E | 3' Flank of R
Used when subcloning an Effector into the 3' domain of an IRES shuttle vector. Compatible cohesive end with PacI in the 5' flank of the IRES GFP.
SalI
5' Flank of the R Domain
Used to release an entire Transcription Unit from vector backbone, or when adding a second Transcription Unit - subcloning your SalI into your first Transcription Unit Linker's XhoI. Compatible cohesive end with XhoI.
ScaI
5' Flank of EL | 3' Flank of E
Used when subcloning an Effector into the 5' domain of an IRES shuttle vector. Compatible cohesive end with EcoRV in the 3' flank of the IRES RFP.
XhoI
5' Flank of the TUL Domain
Used to open the TUL to put a Monogene or a Multi-gene into the TUL (at XhoI and NotI). A compatible cohesive end with SalI
EcoRV
IRES-Only Enzyme
Exclusively used in our IRES shuttle vectors.
PacI
IRES-Only Enzyme
Exclusively used in our IRES shuttle vectors.