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Product Overview
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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.
Pre-indexed Assembled Tn5 Transposomes in a 96-well Plate
Tn5 is also available in a 96-well format: Pre-indexed Assembled Tn5 Transposomes. This breakaway 96-well plate is a flexible format to enable higher throughput ATAC-Seq assays. Each well contains a transposome loaded with a unique i5 and i7 index, enabling direct indexing during tagmentation. By having a unique i7 & i5 Indexed Primer combination in each well, it is easy to perform multiple ATAC-Seq reactions without having to pipette primers separately and keep track of the combinations. The ATAC-Seq Buffer Set is a convenient set of buffers to use for 96 ATAC-Seq reactions with the Pre-indexed Assembled Tn5 Transposomes.
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.
Tn5 Overview
Each lot of our Recombinant Tn5 Transposase is QC tested by SDS-PAGE to show a protein band at ~55 kDa with >90% purity (Figure 1), and by a Tagmentation Test on genomic K562 cell DNA (Figure 2) to confirm that a majority of DNA is observed at 200 – 400 bp with an average sample intensity >1000 by analysis on Agilent TapeStation. See the data.
We use our Recombinant Tn5 Transposase in the Assembled Transposomes Component in the ATAC-Seq Assay Kits, and in our bulk ATAC-Seq Services.
pA-Tn5 Overview
Each lot of our Recombinant pA-Tn5 is QC tested by SDS-PAGE to show a protein band at ~73 kDa with >90% purity (Figure 3), and by a Tagmentation Test on genomic K562 cell DNA (Figure 4) to confirm that a majority of DNA is observed at 200 – 400 bp with an average sample intensity >1000 by analysis on Agilent TapeStation. See the data.
We use our pA-Tn5 in the CUT&Tag-IT® Assembled Transposomes Component in the CUT&Tag-IT® Assay Kits, and in our CUT&Tag-IT® Services.
Pre-indexed Assembled Tn5 Transposomes Overview
It’s easy to perform up to 96 ATAC-Seq reactions with Pre-indexed Assembled Tn5 Transposomes
Pre-indexed Assembled Tn5 Transposomes require no additional loading step, simply add 50,000 to 100,000 prepared cells per well, and incubate with Tagmentation Master Mix for 30 minutes.
During the incubation, unique index combinations are integrated directly into the tagmented DNA. Each well contains a unique combination of i5 and i7 Indexed Primers, simplifying pipetting steps and making it easy to perform multiple reactions without keeping track of the primer combinations one-by-one.
Then, stop the reaction and collect tagmented DNA using SPRI beads and purify the DNA. The libraries are then ready for sequencing on an Illumina® Sequencing System. Universal P5 and P7 Primers are included with the Pre-indexed Assembled Tn5 Transposomes for simple, index-free PCR amplification. Download the protocol and view the plate map for complete details.
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
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
2BFigure 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
4BFigure 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.
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.
Product Documents
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| Name | Format | Cat No. | Price | |
|---|---|---|---|---|
| pA-Tn5 Transposase | 10 µg | 53161 | ¥8,520 | Add to Cart |
| 100 µg | 53162 | Get Quote | ||
| Recombinant Tn5 Transposase protein | 10 µg | 81286 | ¥3,080 | Add to Cart |
| 100 µg | 81284 | Get Quote | ||
| Pre-indexed Assembled Tn5 Transposomes | 1 x 96 rxns | 53152 | ¥17,160 | Add to Cart |
| ATAC-Seq Buffer Set | 1 set | 53153 | ¥7,800 | Add to Cart |
| CUT&Tag-IT® Assay Buffer Set - Cells | 1 set | 53169 | ¥9,100 | Add to Cart |
| CUT&Tag-IT® Assembled pA-Tn5 Transposomes | 16 rxns | 53164 | ¥4,550 | Add to Cart |
| 64 rxns | 53166 | ¥14,560 | Add to Cart |
