24 January 2022

Construction of Strand-seq libraries in open nanoliter arrays

Vincent C.T. Hanlon, Daniel D. Chan, Zeid Hamadeh, Yanni Wang, Carl-Adam Mattsson, Diana C.J. Spierings, Robin J.N. Coope, Peter M. Lansdorp - Cell Reports Methods, 2022


Single-cell Strand-seq generates directional genomic information to study DNA repair, assemble genomes, and map structural variation onto chromosome-length haplotypes. We report a nanoliter-volume, one-pot (OP) Strand-seq library preparation protocol in which reagents are added cumulatively, DNA purification steps are avoided, and enzymes are inactivated with a thermolabile protease. OP-Strand-seq libraries capture 10%–25% of the genome from a single-cell with reduced costs and increased throughput.

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