New England Biolabs is working diligently to ensure we keep our employees and their families safe, while maintaining our business continuity. Despite a "Stay At Home" advisory being put in place in Massachusetts, USA, we are deemed an essential business, and our manufacturing and distribution teams continue to be fully operational. We are continuing to work with our suppliers and distribution partners to ensure uninterrupted access to our products and technical support.
HomeFAQsCan I use Q5U Hot Start High-Fidelity DNA Polymerase for USER® cloning methods?
FAQ: Can I use Q5U Hot Start High-Fidelity DNA Polymerase for USER® cloning methods?
Yes. Q5U Hot Start High-Fidelity DNA Polymerase is a modified Q5 High-Fidelity DNA polymerase which efficiently amplifies uracil-containing templates and primers. For USER cloning methods, target DNA molecules and cloning vector are generated by PCR with 8-12 bases of homology between two fragments. PCR primers start with a 5′ A and contain a single deoxyuracil residue (dU) flanking the 3′ end of the homology region, and can be designed to accommodate multiple-fragment assembly, nucleotide substitutions, insertions and/or deletions. We recommend using the GeneDesign software to design primers for USER junctions. The best results are typically seen when using each primer at a final concentration of 0.5 µM. In comparison to Taq-based enzymes, Q5U Hot Start High-Fidelity DNA Polymerase makes significantly fewer errors, streamlining any cloning protocol (for example, see data below)
You have been idle for more than 20 minutes, for your security you have been logged out. Please sign back in to continue your session.
Your profile has been mapped to an Institution, please sign back for your profile updates to be completed.
Sign in to your NEB account
To save your cart and view previous orders, sign in to your NEB account. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site.