High-throughput single-cell sequencing with reduced amplification bias

Abstract

Provided herein are methods for preparing a sequencing library that includes nucleic acids from a plurality of single cells. In one embodiment, the methods include linear amplification of the nucleicacids. In one embodiment, the sequencing library includes whole genome nucleic acids from the plurality of single cells. In one embodiment, the nucleic acids include three index sequences. Also provided herein are compositions, such as compositions that include the nucleic acids having three index sequences

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS [0002] This application claims the benefit of U.S. Provisional Application Serial No. 62 / 673,023, filed May 17, 2018, and U.S. Provisional Application Serial No. 62 / 821,864, filed March 21, 2019, each of which is hereby incorporated by reference in its entirety. [0003] governmental support [0004] This invention was made with government support under DP1 HG007811 awarded by the National Institutes of Health. The government has certain rights in this invention. technical field [0005] Embodiments of the present disclosure relate to nucleic acid sequencing. In particular, embodiments of the methods and compositions provided herein relate to generating indexed single-cell sequencing libraries and obtaining sequence data therefrom for characterizing rare events, including crossovers and chromosome mis-segregation events. In some embodiments, the method involves resolving cancer heterogeneity at the single-cell level. Bac...

AI Technical Summary

Problems solved by technology

First, most methods require compartmentalization of individual cells, which can limit throughput

Second, most amplification methods are PCR-based and therefore subject to exponential amplification bias

However, this method is low-throughput as it requires the preparation of serial libraries from each single cell (Chen et al., 2017, Science 356:189-194)

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