15-1017CatalogNo.20142001-C1LotNo.

ProductDescrip5on:Recombinantly produced in E. coli, CUTANA pAG-Tn5 forChIC/CUT&Tag is a fusion of Proteins A and G to TransposaseTn5. This construct is useful in performing Cleavage UnderTargets and TagmentaIon (CUT&Tag). The acIve dimer ofTransposase Tn5 is charged with Illumina adapters and readyto be used immediately in CUT&Tag. CUTANA pAG-Tn5 doesnot contain an epitope tag, which makes it compaIble withtag-mediatedCUT&Tag(e.g.FLAG,HA,TY1,etc.).

50reacIonsPackSize

25kDa

37kDa

50kDa

75kDa

100kDa

150kDa

250kDa

20kDa

15kDa

Protein Gel Data: CUTANApAG-Tn5 for ChIC/CUT&Tag(1 µg) was resolved viaSDS-PAGE and stained withCoomassie blue. ThemigraIon and molecularweight of the proteinstandardsareindicated.

Formula5on:

191kDa

Stable for one year at -20°C from date of receipt. Theprotein is not subject to freeze/thaw under thesecondiIons.

(1)Kaya-Okuret.al,Nat.Commun.2019(PMID:31036827)(2)HenikoffandHenikoff,bioRxiv.2020(2020.04.15.043083)

Size Distribu5on of Released Chroma5n: CUT&Tag wasperformed using CUTANA pAG-Tn5 (1:20 diluIon) with100,000 K-562 nuclei. Recovered DNA was directly PCRamplified to produce sequence-ready libraries. AgilentBioanalyzer traces for libraries derived from negaIvecontrol Rabbit IgG (lef; EpiCypher Catalog No. 13-0042,Lot 20036001-52) and H3K27me3 (right; EpiCypherCatalog No. 13-0030, Lot 18303001) are shown. ExcisedDNA is highly enriched for mononucleosomes (peak at~300bpreflects150bpinsertsize).

Mol.Wgt.

StorageandStability:

CUTANA™pAG-Tn5forChIC/CUT&Tag

CUTANA pAG-Tn5 is provided as a 20X stock in storagebuffer (50mM HEPES-KOH pH 7.2, 100mM NaCl, 0.1mMEDTA,1mMDTT,0.1%TritonX-100,50%glycerol).

TransposaseType E.coliExpressedIn

EpitopeTag

Thisproductissufficienttoperform50CUT&TagreacIons.Recommended use: 2.5 µL of the supplied enzyme into a 50 µLCUT&Tag reacIon (20X diluIon). For detailed applicaIons and usesofthisproduct,pleaseseeCUT&Tagprotocolat:epicypher.com/resources/protocolsAdapterSequences:Tn5ME-A:5’-TCGTCGGCAGCGTCAGATGTGTATAAGAGACAG-3’Tn5ME-B:5’-GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAG-3’Tn5MErev:5’-[phos]CTGTCTCTTATACACATCT-3’

None

Applica5onNotes:

References:

Thisproductisforinvitroresearchuseonlyandisnotintendedforuseinhumansoranimals.

EpiCypher,Inc•Phone:855-374-2461•Fax:855-420-6111•www.epicypher.com

CUT&Tag Data: Representa)ve sequencing tracks obtained using CUTANA pAG-Tn5 are shown for two different loci:

a 300 kb close up view of the LAMC3 gene (A) and a wide 5,608 kb window (B). CUT&Tag was performed using

100,000 K-562 nuclei with H3K4me3 an)body (EpiCypher Catalog No. 13-0041, Lot 20083002-42), H3K27me3

an)body (EpiCypher Catalog No. 13-0030, Lot 18303001) and nega)ve control Rabbit IgG (EpiCypher Catalog No. 13

-0042, Lot 20036001-52). Total paired-end read counts were 0.7, 1.1, and 0.9 million reads for H3K4me3,

H3K27me3, and IgG, respec)vely. Images were generated using the Integra)ve Genomics Viewer (IGV, Broad

Ins)tute). CUTANA pAG-Tn5 produced clear peaks with genomic distribu)on profiles consistent with the known

biologicalfunc)onsofH3K4me3andH3K27me3.

Applica'on Notes Addendum: Since CUT&Tag has lower background and is compa)ble with fewer cells compared to ChIP-

seq, it is not recommended to assess fragment size distribu)on using agarose gel or capillary electrophoresis (e.g. AgilentBioanalyzer or TapeSta)on) prior to library prepara)on. This analysis is not indica)ve of the success of a CUT&Tag experiment,

and further the amount of DNA recovered may be below the sensi)vity of detec)on for these approaches. To gauge the

success of a CUT&Tag experiment, assess DNA yield compared to posi)ve (e.g. H3K4me3, H3K27me3) and nega)ve (IgG)

controls, determine fragment size distribu)on of sequence-ready libraries, and evaluate peak structure and expected genome-

widedistribu)oninNGSdata.

Thisproductisforinvitroresearchuseonlyandisnotintendedforuseinhumansoranimals.

EpiCypher,Inc•Phone:855-374-2461•Fax:855-420-6111•www.epicypher.com