CUTANA™ pAG-Tn5 for ChIC/CUT&Tag - 250 Rxns

SKU: 15-1117
Pack Size: 250 reactions
Type:  Transposase Host: E.coli
Mol Wgt: 191 kDa Epitope Tag: None

CUTANA pAG-Tn5 for ChIC/CUT&Tag Description: Recombinantly produced in E. coli, CUTANA pAG-Tn5 for ChIC/CUT&Tag is a fusion of Proteins A and G to Transposase Tn5. This construct is useful in performing Cleavage Under Targets and Tagmentation (CUT&Tag). The active dimer of Transposase Tn5 is charged with Illumina adapters and ready to be used immediately in CUT&Tag. CUTANA pAG-Tn5 does not contain an epitope tag, which makes it compatible with tag-mediated CUT&Tag (e.g. FLAG, HA, TY1, etc.).

CUTANA pAG-Tn5 for ChIC/CUT&Tag Formulation: CUTANA pAG-Tn5 is provided as a 20X stock in 50mM HEPES-KOH pH 7.2, 100mM NaCl, 0.1mM EDTA, 1mM DTT, 0.1% Triton X-100, 50% glycerol.

CUTANA pAG-Tn5 for ChIC/CUT&Tag Storage and Stability: Stable for one year at -20°C from date of receipt. The protein is not subject to freeze/thaw under these conditions.

CUTANA pAG-Tn5 for ChIC/CUT&Tag Application Notes: This product is sufficient to perform 250 CUT&Tag reactions. Recommended use: 2.5 μL of the supplied enzyme into a 50 μL CUT&Tag reaction (20X dilution). For detailed applications and uses of this product, please see CUT&Tag protocol at:
Adapter Sequences:

Application Notes Addendum: Since CUT&Tag has lower background and is compatible with fewer cells compared to ChIP-seq, it is not recommended to assess fragment size distribution using agarose gel or capillary electrophoresis (e.g. Agilent Bioanalyzer or TapeStation) prior to library preparation. This analysis is not indicative of the success of a CUT&Tag experiment, and further the amount of DNA recovered may be below the sensitivity of detection for these approaches. To gauge the success of a CUT&Tag experiment, assess DNA yield compared to positive (e.g. H3K4me3, H3K27me3) and negative (IgG) controls, determine fragment size distribution of sequence-ready libraries, and evaluate peak structure and expected genome-wide distribution in NGS data.

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. 15-1117 Datasheet
View CUTANA CUT&Tag Protocol for Histone PTMs and Transcription Factors. CUTANA<sup>™</sup> CUT&Tag Protocol
View previous CUTANA CUT&Tag Protocol. CUTANA<sup>™</sup> CUT&Tag Protocol
15-1117 Protein Gel Data

Protein Gel Data: CUTANA pAG-Tn5 for ChIC/CUT&Tag (1 µg) was resolved via SDS-PAGE and stained with Coomassie blue. The migration and molecular weight of the protein standards are indicated.
(Click image to enlarge)

15-1117 Size Distribution of Released Chromatin

Size Distribution of Released Chromatin: CUT&Tag was performed using CUTANA pAG-Tn5 (1:20 dilution) with 100,000 K-562 nuclei and recovered DNA was directly PCR amplified to produce sequence-ready libraries. Agilent Bioanalyzer traces for libraries derived from negative control Rabbit IgG (left; EpiCypher Catalog No. 13-0042, Lot 20036001-52) and H3K27me3 (right; EpiCypher Catalog No. 13-0030, Lot 18303001) are shown. Excised DNA is highly enriched for mononucleosomes (peak at ~300 bp reflects 150 bp insert size).
(Click image to enlarge)

15-1117 CUT&Tag Data

CUT&Tag Data: Representative 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 antibody (EpiCypher Catalog No. 13-0041, Lot 20083002-42), H3K27me3 antibody (EpiCypher Catalog No. 13-0030, Lot 18303001) and negative 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, respectively. Images were generated using the Integrative Genomics Viewer (IGV, Broad Institute). CUTANA pAG-Tn5 produced clear peaks with genomic distribution profiles consistent with the known biological functions of H3K4me3 and H3K27me3.
(Click image to enlarge)

Current stock: 0