Preparation of Linked-Read Sequencing Libraries using Haplotagging beads

Kucka, M; Chan, YF

doi:10.17504/protocols.io.5jyl827m8l2w/v1

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Preparation of Linked-Read Sequencing Libraries using Haplotagging beads

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Kucka, M
Chan Group, Friedrich Miescher Laboratory, Max Planck Society;

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Chan, YF
Chan Group, Friedrich Miescher Laboratory, Max Planck Society;

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Citation

Kucka, M., & Chan, Y. (2024). Preparation of Linked-Read Sequencing Libraries using Haplotagging beads. Springer Protocols. doi:10.17504/protocols.io.5jyl827m8l2w/v1.

Cite as: https://hdl.handle.net/21.11116/0000-000F-E3AD-E

Abstract

Haplotagging beads are prepared in form of a 96 well plate where each well contains M280-Streptavidin beads linked with complete and barcoded i5 and i7 Tn5-sequencing adapters. Each well contains 884736 well-specific and barcoded Tn5-adapters assembled with Tn5 transposase. Whole plate then has almost 85 million barcodes (96 x 884736 = 84934656 barcodes), with each bead carrying many copies of a single segmented barcode.
The haplotagging beads are used to prepare short-read linked-read sequencing libraries with both i7 and i5 index being 13bp long. High molecule DNA is tagmented with a single bead and all the reads of that DNA molecule will carry the same barcode combination (one of 85 million).