ATAC libraries were prepared from approximately 50 to 75 thousand of mononuclear cells using a Tagment DNA Enzyme Kit from Illumina (Cat # 20034198) by following the Omni-ATAC protocol (23 (link)). The ATAC libraries were run on 2% agarose gel and gel band corresponding to ~200–600 bp was cut to elute DNA fragments using the MinElute® Gel Extraction Kit (Qiagen), subjected to sequencing at the Genomics Core Facility at Marshall University with Illumina Nextseq2000.
Tagment dna enzyme kit
Manufactured by Illumina
The Tagment DNA Enzyme Kit is a laboratory tool designed to prepare DNA samples for sequencing. It contains the necessary reagents and enzymes to fragment and tag DNA, a process known as 'tagmentation'. This enables the subsequent attachment of adapter sequences, which is a crucial step in the DNA sequencing workflow.
Automatically generated - may contain errors
2 protocols using tagment dna enzyme kit
1
ATAC-seq Library Preparation from Mononuclear Cells
2
Single-cell ATAC-seq of SPI1 mutant RS4;11 cells
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100,000-cell aliquots of SPI1+/+, SPI1+/−, and SPI1−/− RS4;11 cells were harvested in triplicate, washed twice, resuspended in 50 µl cold lysis buffer (10 mM Tris-HCl, pH 7.4, 10 mM NaCl, 3 mM MgCl2, and 0.1% IGEPAL CA-630), and then spun down immediately at 550 ×g for 10 min, 4°C. The supernatant was discarded, and nuclei were immediately resuspended in cold transposition reaction mix from Illumina Tagment DNA Enzyme kit. The transposition reaction was incubated at 37°C for 45 min. Immediately following transposition, 10 µl of 3M Sodium Acetate was added to the DNA, which was then purified using a Qiagen MinElute Kit. Transposed DNA fragments were PCR amplified for 12 cycles using NEBNext High-Fidelity 2× PCR Master Mix with Nextera primers (Illumina). The PCR reaction was cleaned up using AMPureXP beads (Agencourt) and then paired-end sequenced on the Illumina NovaSeq platform (51-bp read length) at the Center for Spatial and Functional Genomics.
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