The impact of ambient contamination on demultiplexing methods for single-nucleus multiome experiments.

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Tác giả: Kevin Abuhanna, Marcus Alvarez, Brunilda Balliu, Jason Ernst, Terence Li, Cuining Liu, Chongyuan Luo, Kathrin Plath, Yu Sun, Noah Zaitlen

Ngôn ngữ: eng

Ký hiệu phân loại: 571.66 +Nucleus

Thông tin xuất bản: United States : Research square , 2025

Mô tả vật lý:

Bộ sưu tập: NCBI

ID: 669145

Sample multiplexing has become an increasingly common design choice in droplet-based single-nucleus multi-omic sequencing experiments to reduce costs and remove technical variation. Genotype-based demultiplexing is one popular class of methods that was originally developed for single-cell RNA-seq, but has not been rigorously benchmarked in other assays, such as snATAC-seq and joint snRNA/snATAC assays, especially in the context of variable ambient RNA/DNA contamination. To address this, we develop ambisim, a genotype-aware read-level simulator that can flexibly control ambient molecule proportions and generate realistic joint snRNA/snATAC data. We use ambisim to evaluate demultiplexing methods across several important parameters: doublet rate, number of multiplexed donors, and coverage levels. Our simulations reveal that methods are variably impacted by ambient contamination in both modalities. We then applied the demultiplexing methods to two joint snRNA/snATAC datasets and found highly variable concordance between methods in both modalities. Finally, we develop a new metric,
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