CUTANA High Fidelity 2X PCR Master MixUser Manual Data sheet
CUTANA High Fidelity 2X PCR Master Mix for CUT&Tag is formulated for non-hot start amplification of next generation sequencing (NGS) libraries from CUT&Tag experiments using CUTANA pAG-Tn5. The master mix requires only the addition of primers and the tagmented chromatin to achieve high fidelity amplification of NGS libraries.
Q5® High Fidelity thermostable DNA Polymerase (3’→5’ exonuclease activity) fused to Sso7d processivity-enhancing domain in a master mix containing 200 μM dNTPs and 4 mM Mg2+ in proprietary buffer.
After completing pAG-Tn5 chromatin tagmentation, add 25 μL of the supplied enzyme into a ~25 μL CUT&Tag reaction (2X dilution). For detailed instructions regarding non-hot start PCR cycling conditions in CUT&Tag, see CUTANA Direct-to-PCR CUT&Tag protocol: epicypher.com/resources/protocols.
NOTE: Not recommended for use with primers or templates containing uracil.
(1) Kaya-Okur et. al, Nat. Commun. 2019 (PMID : 31036827)
(2) Henikoff and Henikoff, bioRxiv. 2020 (2020.04.15.043083)