Illumina library applications for 2x500 bp sequencing on the MiSeq i100

Illumina launched MiSeq i100 Series 25M Reagent Kit (1000 cycles) on October 20, 2025, to enable 2x500 bp sequencing on the MiSeq i100. Illumina has tested modifications to existing library preparation workflows to increase library sizing to leverage these longer read lengths.

Resources for longer insert library preparation will be linked within this article as they become available.

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