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we have determined the amplification cycle number we will now perform sample index PCR and spry select cleanup we have amp Mix B ortex centrifuge briefly and place on ice we have the Dual index plate TS set a we have spry select reagent freshly prepared 80 ethanol but for EB and a 10x magnetic separator finally we have the samples in an eight tube strip choose the appropriate sample index sets to ensure that no sample indices overlap in a multiplexed sequencing run record the 10x sample index name used next we will prepare the amplification Master ranks prepare the amplification Master mix by combining nuclease-free water and amp Mix B ipetnics 10 times shortly before use add 70 microliters amplification Master mix to a tube and an a tube strip for each sample add 25 microliters of each sample from pre-amplification to a separate tube previously aliquated with amplification Master mix add 5 microliters of an individual dual index TS set a to each tube and record the well ID use