RM_JAN22 is being repeated starting at PCR to increase the diversity of MIDs. Since we used two consecutive 16S MID plates for the first PCR, we suspect that this is causing the clusters to be too similar to properly pass through the flow cell. To fix this issue, the new PCR set up only used one MID plate (16S0H616S0H5) and was set up as follows:
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1000/32.25 = 31uL of Pool to Add
100 1000 - uL of Pool to Add = ul of “10 mM Tris 8.5” to Add
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