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==PCR Indexing== Similar to transposon-level indexing, PCR level indexing was also performed with 8 i5 and 12 i7 PCR primers #Add 10 uL of 5 uM i5 primer to its respective well after SDS treatment #Aliquot 30 uL of Nextera PCR Master mix and water into each well from a master mix to create a total PCR volume of 100 uL #For PCR ##72C for 3 min ##98C for 30 sec ##20 cycles ###98C for 30 sec ###63C for 30 sec ###72C for 3 min #Pool all 96 PCRs and purify ##50 uL aliquots pooled ##Assess quality with High-Sensitivity Bioanalyzer
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