The unique enzyme blend of Taq DNA Polymerase and proofreading enzyme combines with the innovative buffers to successfully amplify templates with a GC-content in excess of 70%. The kit includes detailed instructions full of useful tips.
Features & Benefits:
- Efficient amplification up to 40 kb with high yields
- Low error rate and optimized processivity with proofreading enzyme
- Minimal optimization with self-adjusting Mg2+ buffer technology
- Optimized amplification of complex or rare targets
High Yields from Complex Targets up to 40kb
The 5 Prime PCR Extender System combines a powerful polymerase blend with an innovative two buffer system for efﬁcient ampliﬁcation of products ranging from 100 bp up to 40 kb, including GC-rich targets and other difﬁcult templates. The PCR Extender Polymerase Mix is a blend of thermostable DNA polymerases that includes a processivity-enhancing factor. This unique mixture enables extremely high extension rates, maximum proofreading ﬁdelity and high ampliﬁcation efﬁciency, even when using complex templates. This features ensure the optimal ampliﬁcation of PCR products for cloning and mutagenesis.
Fig.1: Amplification of 10-40 kb fragments from Lamda DNA and 4.8-26 kb fragments from human DNA with PCR Extender system using the special Tuning Buffer. Double bands (gDNA) are caused by amplification of the two different alleles
Optimized processivity with a two buffer system for high-ﬁdelity PCR of long targets:
The two buffer systems provided in the 5 Prime PCR Extender System, the Tuning Buffer and the PCR Extender Buffer, provide the unique zwitterionic formulation that improves pH-maintenance at high temperatures (72–94°C). This system reduces pH-driven template degradation to a minimum. The Tuning Buffer is designed for high-ﬁdelity PCR applications and robust ampliﬁcation of genomic targets >20 kb and episomal targets up to 40 kb without organic cosolvents. The PCR Extender Buffer is designed for high ﬁdelity ampliﬁcations of smaller targets ranging from 100 bp to 10 kb genomic DNA and up to 15 kb plasmid or phagemid.
Fig 2: The optimal enzyme & buffer combination provides extremely long fragments of 40 kb.
Storage & Dilution Buffers:
20 mM Tris-HCl pH 8.0 (at 25°C), 100 mM KCl, 0.1 mM EDTA, 1 mM DTT, 50% glycerol, 0.5% Tween 20.
10x Tuning Buffer with Magnesium, 10x High-fidelity Buffer with Magnesium.
Separate Magnesium Solution:
25 mM Mg2+.
Storage at –20°C in a constant-temperature freezer.
- Cat # 2200500: 5 Prime PCR Extender System – 100U
- Cat # 2200520: 5 Prime PCR Extender System – 500U