CUTANA™

CUTANA™ High Fidelity 2X PCR Master Mix

$115.00
SKU: 15-1018
Pack Size: 50 reactions
Type:  Polymerase Host: E.coli
Mol Wgt: N/A Epitope Tag: None


CUTANA™ High Fidelity 2X PCR Master Mix for CUT&Tag Description: 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 (EpiCypher Catalog No. 15-1017 & 15-1117). The master mix requires only the addition of primers and the tagmented chromatin to achieve high fidelity amplification of NGS libraries.

CUTANA™ High Fidelity 2X PCR Master Mix for CUT&Tag Formulation: 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.

CUTANA™ High Fidelity 2X PCR Master Mix for CUT&Tag Storage and Stability: Stable for 6 months at -20°C from date of receipt. The master mix should be thawed and resuspended prior to use.

CUTANA™ High Fidelity 2X PCR Master Mix for CUT&Tag Application Notes: This product is sufficient to perform PCR amplification for 50 CUT&Tag samples.
Recommended use: 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.

Product References:
(1) Kaya-Okur et. al, Nat. Commun. 2019 (PMID : 31036827)
2) Henikoff and Henikoff, bioRxiv. 2020 (2020.04.15.043083)

View technical datasheet for this product. 15-1018 Datasheet
15-1018 CUT&Tag Next Generation Sequencing Libraries

CUT&Tag Next-Generation Sequencing (NGS) Libraries: CUT&Tag DNA from 100,000 K-562 nuclei was directly PCR amplified using CUTANA High Fidelity 2X PCR Master Mix to produce NGS libraries. Tapestation size distribution analysis is shown for CUT&Tag libraries using H3K4me3 (top; EpiCypher Catalog No. 13-0041) and H3K27me3 (bottom; EpiCypher Catalog No. 13-0030) antibodies. The High Fidelity Master Mix successfully amplified NGS libraries predominantly enriched for mononucleosome sized fragments (~300 bp peaks reflect nucleosomes + sequence adapters).
(Click image to enlarge)

15-1018 CUT&Tag NGS Data

CUT&Tag NGS Data:CUT&Tag DNA from the CUTANA High Fidelity 2X PCR Master Mix amplified NGS libraries (above) were subjected to paired-end sequencing using a NextSeq 2000. A 240 bp representative locus is shown for H3K4me3 (top; EpiCypher Catalog No. 13-0041) and H3K27me3 (bottom; EpiCypher Catalog No. 13-0030). NGS libraries amplified using the CUTANA 2X PCR Master Mix faithfully represented the expected genomic distribution of each histone modification. Image was generated using the Integrative Genomics Viewer (IGV, Broad Institute).
(Click image to enlarge)

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