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===Quantify=== #qPCR all 8 samples 1ul each with triplicates {| {{table}} | align="center" style="background:#f0f0f0;"|'''Components''' | align="center" style="background:#f0f0f0;"|'''1X Volume''' | align="center" style="background:#f0f0f0;"|'''25X Volume''' |- | Captured template||1||0 |- | 10uM ISB_CA_AF||0.4||10 |- | 10uM ISB_CA_AR.T2||0.4||10 |- | 2X KAPA SYBG MM||12.5||312.5 |- | H2O||10.7||267.5 |- | Total||25||600 |} *Aliquot 24ul from 24X master mix and add 1ul captured template Program 98C 1min -> (98C 10s -> 52C 20s -> 72C 20s)x30 -> 72C 3min [[File:20170622_Agi15kFeb17_invitrocapture_SplintR5formamide.PNG|350px]][[File:20170622_Agi15kFeb17_invitrocapture_SplintR5formamide_Ct.PNG|250px]] *[[Media:20170622_Agi15kFeb17_invitrocapture_SplintR5formamide.xlsx|Excel Data]] *No separation between DNA, RNA, and NTC samples for SplintR *Sequence results to see if there's no difference really **Stop Ampligase NTC at 28 cycles **Stop others at 19 cycles
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