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  • Run under standard conditions

Results:

16S Pool: 0.48 nM

ITS Pool: 3.61 nM

Sequencing:

  • 1

    0.2 nM 16S Pool:

    100/nM pool concentration= ul Pool to add
  • 100-ul Pool to add=ul of 10 mM Tris to add

  • Add 5 ul 1 nM Pool to 85 42.2 ul 16S Pool + 57.8 ul “10 mM Tris 8.5”

  • 0.2 nM ITS Pool: 5.6 ul ITS Pool + 94.4 ul “10 mM Tris 8.5”

  • 0.1 nM Full Pool: 50 ul from both 0.2 nM Pools

  • Add 50 ul 0.1 nM Pool to 40 ul “10 mM Tris 8.5” and 10 ul 50 pM PhiX

  • Remove iSeq100 Cartridge v2 from large freezer #3 36 hours to 1 week before sequencing and place in refrigerator 5 with room around it for full airflow

  • Remove iSeq 100 i1 Flow Cell from refrigerator 5’s crisper drawer and open white foil pack and allow to equilibrate to RT for 10-15 minutes.

  • Open “iSeq 100 i1 Reagent Cartridge v2”. Turn on iSeq100, sbsuser password: “GenomeTechnologies”.

  • Click on “Sequence”. Watch Video. Do what video tells you to do. Follow on screen instructions until run starts.

  • Results located: Data/SequencingRuns/”folder with applicable date”/Alignment_1/Fastq/*.fastq.gz

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