Recombinant Transposase Enzymes

Product Overview

Pre-indexed Assembled Tn5 Transposomes

ATAC-Seq Buffer Set

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The recombinant Tn5 and pA-Tn5 enzymes are useful for many applications, including ATAC-Seq, CUT&Tag assays, Tagmentation, ChIPmentation, and more.

Essentials for high-throughput ATAC-Seq

Flexible high-throughput ATAC-Seq reagents—Pre-indexed Tn5 Transposomes, Buffer Set, and Q5 Polymerase & Library Purification Kit—available individually or together as the High-Throughput ATAC-Seq Express solution.

Pre-indexed Assembled Tn5 Transposomes are available in a flexible 96-well breakaway plate format designed for high-throughput ATAC-Seq. Each well contains a transposome preloaded with unique i5 and i7 indexes, enabling direct indexing during tagmentation and eliminating the need to pipette primers or track index combinations.

The ATAC-Seq Buffer Set provides all required buffers for 96 reactions, while the Q5 Polymerase and Library Purification Kit includes Q5 Polymerase, silica columns, and SPRI beads for efficient library amplification and cleanup.

Together, these components deliver a complete High-Throughput ATAC-Seq Express solution (Cat. No. 53158) for streamlined preparation of up to 96 ATAC-Seq libraries.

Loaded or unloaded pA-Tn5 and CUT&Tag-IT Assay Buffer Set

pA-Tn5 is available unloaded in 10 µg (Cat. No. 53161) and 100 µg (Cat. No. 53162) sizes to be assembled with adapters of your choice, or pA-Tn5 is available loaded in CUT&Tag-IT Assembled pA-Tn5 Transposomoes in 16 (Cat. No. 53164) or 64 (Cat. No. 53166) reactions. Assembled pA-Tn5 Transpsomes can be used in CUT&Tag assays by following the protocol for CUT&Tag-IT Assay Kit Anti-Rabbit (Cat. No. 53160) or Anti-Mouse (Cat. No. 53165) and using the CUT&Tag-IT Assay Buffer Set – Cells (Cat. No. 53169). The CUT&Tag-IT Assay Buffer Set – Cells provides the essential reagents for 16 CUT&Tag-IT Assay reactions.

Product Contents

Pre-indexed and Assembled Tn5 Transposomes (Cat. No. 53152) includes:

• Pre-indexed and Assembled Tn5 Transposomes, store at -20°C
• P5 Primer (25 µM), store at -20°C
• P7 Primer (25 µM), store at -20°C

ATAC-Seq Buffer Set (Cat. No. 53153) includes:

• ATAC Lysis Buffer, store at RT
• 2X Tagmentation Buffer, store at -20°C
• 1% Digitonin, store at -20°C

CUT&Tag-IT^® Assay Buffer Set (Cat. No. 53169) includes:

• Tagmentation Buffer, store at -20°C or 4°C
• Dig-Wash Buffer, store at 4°C
• Dig-300 Buffer, store at 4°C
• Antibody Buffer, store at 4°C
• 1X Binding Buffer, store at 4°C
• 1X Wash Buffer, store at 4°C

Q5 Polymerase and Library Purification Kit (Cat. No. 53159) includes:

• 5X Q5 Reaction Buffer, store at -20°C
• dNTP Mix (10 mM each), store at -20°C
• Q5 DNA Polymerase, store at -20°C
• SPRI Beads, store at 4°C
• Silica Beads, store at 4°C

Product Data

Recombinant Transposase Enzymes Data

Tn5 SDS-PAGE

Figure 1: 12.5% SDS PAGE Gel of Recombinant Tn5 Transposase Protein (MW 53kDa).

Tn5 Tagmentation Test

2A

2B

Figure 2: Typical tagmentation profile of K562 genomic DNA after incubation with the Recombinant Tn5 Transposase.
50 ng of K562 genomic DNA was incubated for one hour with loaded Tn5 Transposase and ATAC Tagmentation Buffer. AMPure bead clean up was performed on the samples and was followed by Q5 PCR amplification. After another AMPure bead clean up, samples were analyzed for fragment size (gel image 2A) and electropherogram (2B) on the Agilent TapeStation.

pA-Tn5 SDS PAGE Gel

Figure 3: 12.5% SDS PAGE Gel of Recombinant pA-Tn5 Transposase Protein (MW 72kDa).

pA-Tn5 Tagmentation Test

4A

4B

Figure 4: Typical tagmentation profile of K562 genomic DNA after incubation with the Recombinant pA-Tn5 Transposase.
50 ng of K562 genomic DNA was incubated for one hour with loaded pA-Tn5 Transposase and ATAC Tagmentation Buffer. AMPure bead clean up was performed on the samples and was followed by Q5 PCR amplification. After another AMPure bead clean up, samples were analyzed for fragment size (gel image 4A) and electropherogram (4B) on the Agilent TapeStation.

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Pre-indexed Assembled Tn5 Transposomes Data

Figure 5: ATAC-Seq Results are Consistent with Pre-indexed Assembled Tn5 Transposomes and Standard Tn5 Transposomes.
50,000 fresh K562 cells were assayed in ATAC-Seq with either Active Motif’s Pre-indexed Assembled Tn5 Transposomes (top four tracks) or standard, non-indexed Tn5 Assembled Transposomes (bottom track). ATAC-Seq results were consistent between both methods. Browser tracks are shown at Chr6:159,668,373-159,821,466.

Figure 6: i5 and i7 Indexed Primer Combinations Deliver Consistent Results.
ATAC-Seq was performed on 2 replicates of 50,000 fresh K562 cells were assayed with Pre-indexed Assembled Tn5 Transposomes. i5 and i7 Indexed Primer combinations were optimized to yield consistent results. A subset of data is shown. The number of filtered peaks and FRIP score percentages were consistent across all index combinations.

Figure 7: Pre-indexed Assembled Tn5 Transposomes are Compatible Across Different Cell Lines.
100,000 cryopreserved HEK293, K562, HeLa, and GM12878 cells were assayed in ATAC-Seq with Pre-indexed Assembled Tn5 Transposomes resulting in consistent peak intensity as shown in this IGV browser view at Chr19:41,855,000-41,890,000.

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Product Documents

You might also be interested in:

• ATAC-Seq Kits
• CUT&Tag-IT Assay Kits
• End-to-end ATAC-Seq Services
• Single-cell ATAC-Seq Service
• CUT&Tag Services