CUTANA™

CUTANA™ High Fidelity 2X PCR Master Mix

$125.00
SKU: 15-1018
Pack Size: 50 reactions
  • Type:  Polymerase
  • Mol. Wgt.:  N/A
  • Expressed In:  E. coli
  • Epitope Tag:  None

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 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.

Validation Data

Figure 1: CUT&Tag Next-Generation Sequencing (NGS) libraries
CUT&Tag DNA from 100,000 K562 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 IgG negative control (EpiCypher 13-0042) or H3K27me3 (EpiCypher 13-0030) and anti-mouse secondary (EpiCypher 13-0048) antibodies. The High Fidelity Master Mix successfully amplified NGS libraries predominantly enriched for mononucleosome sized fragments (~300 bp peaks reflect nucleosomes + sequence adapters).

Figure 2: CUT&Tag NGS data
CUT&Tag DNA from the CUTANA™ High Fidelity 2X PCR Master Mix amplified NGS libraries were subjected to paired-end sequencing using an Illumina NextSeq2000. A 240 bp representative locus is shown for IgG (EpiCypher 13-0042) or H3K27me3 (EpiCypher 13-0030). NGS libraries amplified using the CUTANA™ 2X PCR Master Mix represented the expected genomic distribution of H3K27me3. Image was generated using the Integrative Genomics Viewer (IGV, Broad Institute).

Technical Information

Formulation
A high fidelity thermostable DNA polymerase with 3'→5' exonuclease activity fused to a processivity-enhancing domain in a 2x master mix containing 200 µM dNTPs and 4 mM Mg2+ in a proprietary buffer.
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.

Application Notes

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.

NOTE: Not recommended for use with primers or templates containing uracil.

References

Background References:
[1] Kaya-Okur et al. Nat. Commun. (2019). PMID: 31036827
[2] Henikoff et al. Elife. (2020). PMID: 33191916

Product References:

Documents & Resources

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