Enzymes used for Bird of Prey (BoP)
We are proud to be working with New England Biolabs (NEB) to provide OspreyBio customers a discounted rate on a pack of ten enzymes that will be ready for purchase on their site in early 2024. In the meantime, below are the ten enzymes required to fully unlock the power of Bird of Prey and your CloneCards. For a more detailed tutorial, please go to our Bird of Prey Academy Page.
While we recommend purchasing all enzymes at first, some will likely be used more than others depending on what you want to achieve.
Below, we describe the use of each enzyme as well as provide a link to NEB's site for each by clicking on the names.
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.
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.
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.
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.
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.
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.
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.
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
IRES-Only
Enzymes
EcoRV - PacI