HomeFAQsWhy is there a 55°C, 5 min heat step at the end of the assembly reaction?
FAQ: Why is there a 55°C, 5 min heat step at the end of the assembly reaction?
The final incubation step at 55°C favors BsaI cutting, in the absence of DNA
ligation. Digesting any plasmid still present in the assembly reactions reduces
background. Golden Gate Assembly can tolerate one internal BsaI site in one
fragment; in this case omit the 55°C step and screen more colonies as the
background will be higher.
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.