AccuNext ATAC DNA Library Building Kit (Illumina)

This is an ATAC-seq library construction kit developed for Illumina's high-throughput sequencing platform for 10³~10⁵  Sample building for a starting volume of one cell.ATAC-seq (Assay for Transposase Accessible Chromatin with high-throughput sequencing) is a high-throughput sequencing technology mediated by transposase for rapid detection of open regions of chromatin on a genome-wide scale; it can reveal the association between chromatin accessibility and gene expression regulation, providing key clues for epigenetic studies; it is applicable to epigenetic studies such as chromosome openness mapping, embryonic development, prediction of potential markers of diseases, tumorigenesis, etc.
This product uses Tn5 transposon containing part of illumina sequencing junction, combined with chromatin open region, to cut naked DNA and add sequencing junction at the same time, and then obtain sequencing library after PCR amplification, which has the advantages of wide range of cellular starting range, short experimental period, high signal-to-noise ratio, and wide range of applications.
The reaction system of this product has been carefully optimized, all reagents in the experiment should be used as provided in this product, it is not recommended to change any reaction components or replace the components in this product with other equivalent products in order to avoid obtaining bad experimental results. If replacement is needed, please verify it first.

 

1. For illumina sequencing platform: integrates Index and illumina connectors and constructs DNA libraries that can be directly sequenced by NGS.
2. High throughput: ATAC-seq is capable of analyzing a large number of samples in a short period of time, which is suitable for large-scale genomic research.
3. Low Cell Requirement: ATAC-seq requires a lower number of cells than traditional methods and can be performed from as low as 1000 cells.
4. Wide range of cellular inputs: applicable for 10³~10⁵ Sample building for a starting volume of cells.
5. High Repeatability: The experimental results have a high degree of repeatability and consistency.
6. High signal-to-noise ratio: TSS is well enriched and has a high signal-to-noise ratio.

 

Package 3-1 components are as follows (stored at -80°C):

individual parts making up a compound	AG12566	AG12567
AccuNext Tn5 Transposome	16 μl	50 μl

The Package 3-2 components are as follows (stored at 4°C):

individual parts making up a compound	AG12566	AG12567
DNA Clean Beads	950 μl	2.9 ml
Stop Buffer	80 μl	240 μl

Package 3-3 components are as follows (stored at -20°C):

individual parts making up a compound	AG12566	AG12567
Lysis Buffer	200 μl	600 μl
Wash Buffer Ⅰ	1 ml X 2 pcs	1.5 ml X 4 pcs
Wash Buffer II	1.6 ml	1.6 ml X 3 pcs
Tn5 Reaction Buffer	200 μl	600 μl
1% Digitonin	4 μl	12 μl
AccuNext PCR Mix Ⅱ	80 μl	240 μl
Proteinase K (20 mg / ml )	4 μl	12 μl
Nuclease free water	1 ml	1 ml

Attention:AccuNext Transposome building adaptor primers (Illumina) (Code No. AG12532 / AG12533 / AG12534) are required for the experiment, but are not configured in this product and need to be purchased separately.AccuNext Information on the transposome building adaptor primers (Illumina) is provided in Appendix 1.

 

Storage Temperature: Package 3-1 -80℃ for storage
Package 3-2 4°C Storage (avoid freezing)
Package 3-3 -20°C Storage
Transportation Temperature: Package 3-1 Dry Ice Transportation
Package 3-2 Ice Pack Storage
Package 3-3 Dry Ice Transportation or -20℃ Ice Pack Transportation

1. The library of GM12878 Cells was constructed using this product, and the cell numbers were 100000 (cycle number 13), 50000 (cycle number 13) and 1000 (cycle number 19). The library was tested by agarose gel electrophoresis and Agilent 2100 Bioanalyzer, and the electrophoresis results were as follows:

The Agilent 2100 Bioanalyzer assay results were as follows:

Figure 2. Agilent 2100 Bioanalyzer library length assay for different cell volume

The above figure shows the distribution of the libraries of 100000 Cells, 50000 Cells and 1000 Cells, and it can be seen that the distribution of nucleosomes is obvious, the first peak is around 170~200 bp, which is the nucleosome Free model; the second peak appeared later is the one nucleosome model; the third peak is the two nucleosome model.