METHOD

Designing of custom barcodes for sequencing on the MGI platform

Shmitko AO, Bulusheva IA, Vasiliadis YuA, Suchalko ON, Syrko DS, Belova VA, Pavlova AS, Korostin DO
About authors

Pirogov Russian National Research Medical University

Correspondence should be addressed: Аnna O. Shmitko
Ostrovityanova, 1/1, Мoscow, 117997, Russia; moc.liamg@79imhsanna

About paper

Funding: this research was funded by the grant №075-15-2019-1789 from the Ministry of Science and Higher Education of the Russian Federation allocated to the Center for Precision Genome Editing and Genetic Technologies for Biomedicine.

Author contribution: Shmitko AO — study planning, data collection, writing Original Draft Preparation; Bulusheva IA — methodology, data analysis, writing Original Draft Preparation; Vasiliadis IA, Suchalko ON, Syrko DS — data analysis, software, visualization; Belova VA — study planning, manuscript review and editing; Pavlova AS — data curation, data analysis, software; Korostin DO — conceptualization, supervision, methodology, manuscript review and editing.

Received: 2024-08-15 Accepted: 2024-09-17 Published online: 2024-10-10
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The MGI (MGI Tech Co. Ltd., China) next-generation sequencing platform, including the DNBSEQ-G50, -G400, and -T7 sequencers, is being actively adopted in research. Despite its widespread adoption, challenges persist in the form of limitations associated with the manufacturer's provided barcode set for library preparation. These limitations include constraints on the number of samples that can be concurrently sequenced, compatibility issues with barcodes from diverse or incomplete sets, and restrictions on the sample ratio. Purpose: to develop a universal method that allows sequencing of up to 252 samples simultaneously on a single sequencer lane, while eliminating barcode-related limitations. We proposed a “quad method” that provides 4 or 4n+2 equilibration of barcodes. This paper also delves into its comprehensive analysis, verification procedures, seamless integration into the sequencing process and validation of the method on the DNBSEQ G-400 platform. The quad method showed efficiency and reliability, allowing sequencing of up to 252 samples simultaneously without compromising data quality. The proposed method optimizes library preparation and improves the flexibility of sequencing on the MGI platform.

Keywords: NGS, DNBSEQ, MGI, BGI, custom barcodes, custom indexes, custom adapters

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