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Run sample on iSeq:

Dilute to 1 nM based off qPCR results. qPCR results are in pM, but 1:1000 dilution used. The results are effectively in nM for pool.

  • 1000/Results = ul of Pool to Add

  • 1000 - ul of Pool to Add = ul of “10 mM Tris 8.5” to Add

Dilute 1 nM full pool to loading concentration of 50 pM:

  • Add 5 ul 1 nM Pool to 85 ul “10 mM Tris 8.5” and 10 ul 50 pM PhiX

  • 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

  • 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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