Active Motif的ATAC-Seq试剂盒提供了与Illumina®平台兼容的ATAC-Seq文库制备试剂，可以从20 – 30 mg组织或50,000 – 100,000个细胞中生成16种不同的ATAC-Seq文库。经过优化的方案将指导您完成样品制备、DNA片段标记和文库构建的步骤，构建可在单个测序反应中进行多重测序的文库。
- ATAC Lysis Buffer, store at RT
- Assembled Transposomes, store at -20°C
- 2X Tagmentation Buffer, store at -20°C
- 1X PBS, store at -20°C or RT
- 10X PBS, store at RT
- 10% Tween 20, store at RT
- 0.5% Digitonin, store at -20°C
- DNA Purification Columns, store at RT
- DNA Purification Binding Buffer, store at RT
- DNA Purification Wash Buffer, store at RT
- DNA Purification Elution Buffer, store at RT
- 3 M Sodium Acetate, store at RT
- 10 mM dNTPs, store at -20°C
- 5X Q5 Buffer, store at -20°C
- Q5 High-Fidelity DNA Polymerase, store at -20°C
- i7 Indexed Primer 1, store at -20°C
- i7 Indexed Primer 2, store at -20°C
- i7 Indexed Primer 3, store at -20°C
- i7 Indexed Primer 4, store at -20°C
- i5 Indexed Primer 1, store at -20°C
- i5 Indexed Primer 2, store at -20°C
- i5 Indexed Primer 3, store at -20°C
- i5 Indexed Primer 4, store at -20°C
- SPRI Beads, store at 4°C
ATAC-Seq Kit FAQs
Is this kit the same as the original Buenrostro et al Nature 2013 paper?
This kit is based off a combination of the original Buenrostro paper and the Corces et al Nature 2017 paper, and we have optimized the buffers for a more robust and efficient assay.
What is the source of the recombinant Tn5 enzyme in this kit?
The recombinant Tn5 enzyme in this kit is a hyperactive mutant that we cloned, expressed, purified, and test in-house. The Assembled Transposomes containing this enzyme are provided pre-loaded with the next-gen sequencing adapters, so it is ready-to-use without any optimization or other preparation required.
What is the minimum number of cells or tissue per reaction?
50,000 cells or 20 mg tissue.
Can frozen tissue and cells be used?
Yes, but the samples must be high quality to preserve viability. Cells should be cryopreserved in a controlled-rate freeze with media formulated to protect against ice crystal formation and subsequent cell damage, and tissue should be flash frozen at -80°C.
Can nuclei be frozen for later use in ATAC-Seq?
Yes, extracted nuclei can be cryopreserved and frozen at -80°C for later use. Nuclei can also be pooled together to increase the sample size.
Can FACS-sorted cells be used?
It is possible, but cells could be damaged during the FACS-sorting process. For good ATAC-Seq libraries, cells must be viable.
Do you have a recommendation for a 40 micron mesh strainer for tissue samples?
Yes, we recommend the Falcon 40 micron cell strainer (Falcon catalog no. 352340).
I don’t have a thermomixer. Can I still use the kit?
Using a thermomixer is recommended. However, we have tested this and incubated the tagmentation reaction without the use of a mixer at 37°C in a thermal cycler with a heated lid and the reaction was successful.
Can all the reagents and kit components, including master mixes and enzymes, thaw out and be prepared at room temperature?
Everything except the Assembled Transposomes and Q5 DNA polymerase can be thawed at room temperature. The Assembled Transposomes and Q5 DNA Polymerase are formulated with glycerol and will not need thawing, just put these on ice. The other components can be frozen again and thawed again for future reactions, so not all 16 reactions need to be done simultaneously.
What is the expected final library size, and what should the library traces look like on a fragment analyzer?
How many sequencing reads are needed per sample?
Typically 20M reads.