NEB will be closed Thursday, November 23, 2017 and Friday, November 24, 2017. Orders placed will process Monday, November 27, 2017 for delivery on Tuesday, November 28,2017.
HomeFAQsWhat should I use for an annealing temperature with the Q5® Site-Directed Mutagenesis Kit?
FAQ: What should I use for an annealing temperature with the Q5® Site-Directed Mutagenesis Kit?
We recommend using the NEB online primer design tool NEBaseChanger to provide an optimized annealing temperature for mutagenic primer pairs. For predesigned back-to-back primer sets, a Tm+3 rule can be applied but optimization may be necessary. Note that mismatches in the mutagenic primer are not accounted for using standard Tm calculations and the optimal annealing temperature for Q5 High Fidelity Hot Start DNA Polymerase may differ significantly from that of Taq-based polymerases.
For primer pairs with annealing temperatures close to 72°C, a two-step cycling protocol without a separate annealing step can be used.
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.
Sign in to your NEB account
Any items placed in your cart as a guest will be lost upon sign in or leaving the site.