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====Off chip==== Prep * Add 1.2 uL 50X SYBR Green I to 120 uL NPM Nextera XT *Same as standard protocol, but add 15.5 uL NPM/SYBR Green I mix and run on qPCR machine (final volume 50.5 uL) 0.1X Tn5-059 *5 uL total **3 ul template **1 ul SSB **1 ul 1X Tn5-059 *Incubate 55 C for 5 min, transfer to cold block *Add 1 ul 0.2 mg/ml Qiagen Protease *Incubate 50 C for 15 min, followed by heat inactivation at 70 C for 20 min *Add 19 uL H2O to bring volume up from 6 uL to 25 uL *Add 5 uL each index (N7xx/S5xx) *Add 15.5 uL NPM/SYBR Green I mix qPCR protocol *72 C for 3 min *95 C for 1 min *25 cycles (but prepare to stop early) **95 C for 15 sec **55 C for 30 sec **72 C for 30 sec, acquisition **72 C for 15 sec *72 C for 5 min
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