4.6 Article

Improving the efficiency of single-cell genome sequencing based on overlapping pooling strategy and CNV analysis

Journal

ROYAL SOCIETY OPEN SCIENCE
Volume 9, Issue 1, Pages -

Publisher

ROYAL SOC
DOI: 10.1098/rsos.211330

Keywords

single-cell sequencing; copy number variation; intra-tumour heterogeneity; overlapping pooling; branch and bound

Funding

  1. National Natural Science Foundation of China [61971125]
  2. Six Talent Peaks Project of Jiangsu Province [2019-SWYY-004]

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This study presents an efficient method for analyzing copy number variation (CNV) in single cells using overlapping pooling and branch and bound algorithm. By overlapping pooling, the number of required libraries is reduced, resulting in lower costs. Computer simulations showed high accuracy in classifying cells based on CNV pattern, with significantly fewer sequencing libraries required. This method is expected to improve the efficiency of single-cell genome sequencing.
Single-cell genome sequencing has become a useful tool in medicine and biology studies. However, an independent library is required for each cell in single-cell genome sequencing, so that the cost grows with the number of cells. In this study, we report a study which efficiently analyses single-cell copy number variation (CNV) using overlapping pooling strategy and branch and bound (B&B) algorithm. Single cells were overlapped pooled before sequencing, and later were assorted into specific types by estimating their CNV patterns by B&B algorithm. Instead of constructing libraries for each cell, a library is required only for each pool. As the number of pools is smaller than the cells, fewer libraries are required, which means lower cost. Through computer simulations, we overlapped pooled 80 cells into 40 or 27 pools and classified them into cell types based on CNV pattern. The results showed that 84% cells in 40 pools and 76.5% cells in 27 pools were correctly classified on average, while only half or one-third of the sequencing libraries were required. Combining with traditional approaches, our method is expected to significantly improve the efficiency of single-cell genome sequencing.

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