• 10 x 1.25 mL Phusion 预混液(含 HF 缓冲液)(2X)(在最终反应中提供 1.5 mM MgCl2) • 2 x 500 μL 100% DMSO
储存在 -20°C 下。
常见问题解答 (FAQ)
Do Phusion DNA Polymerases add the non-template dependent 3'-A overhang?
Phusion DNA Polymerases generate blunt end products; therefore, blunt end cloning is recommended. If TA cloning is required, it can be performed by adding A overhangs to the blunt PCR product with e.g. Taq DNA Polymerase (Cat. No. EP0401). However, before adding the overhangs it is very important to remove all the Phusion DNA Polymerase by purifying the PCR product carefully, as the proofreading activity in Phusion DNA Polymerase is very strong. Any remaining Phusion DNA Polymerase will degrade the A overhangs, thus creating blunt ends again.
Can Phusion DNA Polymerases extend at 1 second/kb?
Yes it is possible, especially when amplifying smaller amplicons. Processivity of Phusion DNA Polymerases is 10 times that of Pfu. We recommend extension times of 15 s/kb for Phusion Flash PCR Master Mix. 15 s/kb is a conservative value that we can promise to work with almost any amplicon. In many cases, significantly shorter extension times (0-5 s/kb) can be used without compromising the yield. What separates Phusion Flash DNA Polymerase from other fast polymerases is that all steps in the PCR protocol can be shortened, including annealing and denaturation. This results in extremely fast protocols as compared with any other polymerase.
Can protocols optimized for Phusion DNA Polymerase be directly applied to Phusion Hot Start II DNA Polymerase?
Yes, protocols optimized for Phusion DNA Polymerase can be applied to Phusion Hot Start II DNA Polymerase reactions.
Do you have any ready-to-use 2X Master Mix as an alternative to Phusion High-Fidelity PCR Kit (Cat. No. F553L)?
We offer Phusion High-Fidelity PCR Master Mix with HF Buffer (Cat. No. F531L) in a 2X Master Mix format. For GC-rich templates, the Phusion High-Fidelity PCR Master Mix with GC Buffer (Cat. No. F532L) can be used.