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New England Biolabs: How to Amplify GC Rich DNA?

24 Dec 2013
New England Biolabs: How to Amplify GC Rich DNA?

Watch this video from New England Biolabs to learn how OneTaq and Q5 DNA polymerases optimize the amplification of AT rich and GC rich DNA targets respectively.

OneTaq® DNA Polymerase

New England Biolabs Inc.

OneTaq DNA Polymerase is an optimized blend of Taq and Deep VentR™ DNA polymerases for use with routine and difficult PCR experiments. The 3´→ 5´ exonuclease activity of Deep Vent DNA Polymerase increases the fidelity and robust amplification of Taq DNA Polymerase. The OneTaq Reaction Buffers and High GC Enhancer have been formulated for robust yields with minimal optimization, regardless of a template’s GC content.OneTaq DNA Polymerase is supplied with two 5X buffers: (Standard and GC), as well as a High GC Enhancer solution. For most routine and/or AT-rich amplicons (Lambda, etc.) or complex amplicons with up to ~65% GC content, OneTaq Standard Reaction Buffer provides robust amplification. For GC-rich amplicons, the OneTaq GC Reaction Buffer can improve both performance and yield. For particularly high GC or difficult amplicons, the OneTaq High GC Enhancer can be added at a final concentration of 10–20% to reactions containing OneTaq GC Reaction Buffer.OneTaq® DNA Polymerase Features: Ideal for routine, AT- and GC-rich PCR Applications: High Sensitivity PCR High Throughput PCR Routine PCR GC-rich PCR AT-rich PCR Colony PCR Long PCR (up to ~6 kb genomic)

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Q5® High-Fidelity DNA Polymerase

New England Biolabs Inc.

Q5® High-Fidelity DNA Polymerase is a high-fidelity, thermostable DNA polymerase with 3´→ 5´ exonuclease activity, fused to a processivity-enhancing Sso7d domain to support robust DNA amplification. With an error rate > 100-fold lower than that of Taq DNA Polymerase and 12-fold lower than that of Pyrococcus furiosus (Pfu) DNA Polymerase, Q5 High-Fidelity DNA Polymerase is ideal for cloning and can be used for long or difficult amplicons.Q5 High-Fidelity DNA Polymerase is supplied with an optimized buffer system that allows robust amplification regardless of GC content. The 5X Q5 Reaction Buffer contains 2 mM MgCl2 at final (1X) reaction concentrations and is recommended for most routine applications.For GC-rich targets (≥ 65% GC), amplification can be improved by the addition of the 5X Q5 High GC Enhancer. Q5 High-Fidelity DNA Polymerase is unlike typical, lower fidelity PCR enzymes.Q5® High-Fidelity DNA Polymerase Applications: High-fidelity PCR Cloning Long or Difficult Amplification High-throughput PCR

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New England Biolabs: How to Amplify GC Rich DNA?