New England Biolabs, Inc., 240 County Road, Ipswich, MA, 01938, USA.
Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, 450 Brookline Avenue, Boston, MA, 02215-5450, USA.
Clin Epigenetics. 2020 Sep 22;12(1):143. doi: 10.1186/s13148-020-00921-6.
Accessible chromatin plays a central role in gene expression and chromatin architecture. Current accessible chromatin approaches depend on limited digestion/cutting and pasting adaptors at the accessible DNA, thus requiring additional materials and time for optimization. Universal NicE-seq (UniNicE-seq) is an improved accessible chromatin profiling method that negates the optimization step and is suited to a variety of mammalian cells and tissues. Addition of 5-methyldeoxycytidine triphosphate during accessible chromatin labeling and an on-bead library making step substantially improved the signal to noise ratio while protecting the accessible regions from repeated nicking in cell lines, mouse T cells, mouse kidney, and human frozen tissue sections. We also demonstrate one tube UniNicE-seq for the FFPE tissue section for direct NGS library preparation without sonication and DNA purification steps. These refinements allowed reliable mapping of accessible chromatin for high-resolution genomic feature studies.
可及染色质在基因表达和染色质结构中起着核心作用。目前的可及染色质方法依赖于有限的 DNA 可及性消化/切割和接头连接,因此需要额外的材料和时间进行优化。通用 NicE-seq(UniNicE-seq)是一种改进的可及染色质分析方法,它省去了优化步骤,适用于各种哺乳动物细胞和组织。在可及染色质标记和珠上文库制备步骤中添加 5-甲基脱氧胞苷三磷酸,同时保护可及区域免受细胞系、小鼠 T 细胞、小鼠肾脏和人冷冻组织切片中重复的缺口,大大提高了信号与噪声比。我们还展示了一种用于 FFPE 组织切片的单管 UniNicE-seq,无需超声处理和 DNA 纯化步骤,即可直接进行 NGS 文库制备。这些改进使得对高分辨率基因组特征研究的可及染色质进行可靠的作图成为可能。