HomeFAQsWhen using a polymerase that doesn't contain a 3'-5' exonuclease activity (such as Taq DNA Polymerase) to amplify fragments to be used in a Gibson Assembly reaction, should I be concerned about the potential 3' mismatch generated by the addition of a non-templated nucleotide?
FAQ: When using a polymerase that doesn't contain a 3'-5' exonuclease activity (such as Taq DNA Polymerase) to amplify fragments to be used in a Gibson Assembly reaction, should I be concerned about the potential 3' mismatch generated by the addition of a non-templated nucleotide?
This type of 3' mismatch can affect the efficiency of the assembly, especially when attempting to assemble a high number of fragments (4-6). In those cases, it may be necessary to screen more colonies to obtain the correctly assembled clone. To prevent this, we recommend the use of polymerases that produce blunt-ended amplicons, such as Q5 High-Fidelity DNA Polymerase .
Choose your country
North America
Europe
Asia-Pacific
Session Expired
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.
Institution Changed
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.
