Mze N. P., Beye M., Kacel I., Tola R., Basco Leonardo, Bogreau H., Colson P., Fournier P. E. (2022). Simultaneous SARS-CoV-2 genome sequencing of 384 samples on an Illumina MiSeq instrument through protocol optimization. Genes, 13 (9), 1648 [9 p.].
Titre du document
Simultaneous SARS-CoV-2 genome sequencing of 384 samples on an Illumina MiSeq instrument through protocol optimization
Mze N. P., Beye M., Kacel I., Tola R., Basco Leonardo, Bogreau H., Colson P., Fournier P. E.
Source
Genes, 2022,
13 (9), 1648 [9 p.]
In the present study, we propose a high-throughput sequencing protocol using aNextera XT Library DNA kit on an Illumina MiSeq instrument. We made major modifications to this library preparation in order to multiplex 384 samples in a single Illumina flow cell. To validate our protocol, we compared the sequences obtained with the modified Illumina protocol to those obtained with the GridION Nanopore protocol. For the modified Illumina protocol, our results showed that 94.9% (357/376) of the sequences were interpretable, with a viral genome coverage between 50.5% and 99.9% and an average depth of 421x. For the GridION Nanopore protocol, 94.6% (356/376) of the sequences were interpretable, with a viral genome coverage between 7.0% and 98.6% and an average depth of 2123x. The modified Illumina protocol allows for gaining EUR 4744 and returning results of 384 samples in 53.5 h versus four times 55.5 h with the standard Illumina protocol. Our modified MiSeq protocol yields similar genome sequence data as the GridION Nanopore protocol and has the advantage of being able to handle four times more samples simultaneously and hence is much less expensive.
Plan de classement
Sciences fondamentales / Techniques d'analyse et de recherche [020]
;
Entomologie médicale / Parasitologie / Virologie [052]
