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===Probe Expansion PCR=== *Expansion PCR in 200ul total: 2 tubes of 100ul for 0bp gap probes with AP1V6 primers {| {{table}} | align="center" style="background:#f0f0f0;"|'''''' | align="center" style="background:#f0f0f0;"|'''0 gap probes (ul)''' |- | Agi26k (20nM)||10 |- | 2x Kapa SYBG qPCR MM||100 |- | 100uM pAP1V6U||0.8 |- | 100uM AP2V6||0.8 |- | H2O||88.4 |- | Total||200 |- | |} *Mixed thoroughly and then split into two tubes 94C 2min -> (94C 30sec -> 54C 45sec -> 72C 45sec) x 12 -> 72C 3min -> 15C hold [[File:20140522_Agi26k0gapExpansionPCR.JPG| 450px]] *Purified with 2 Qiagen columns *Eluted with 40ul each tube and combined into two tubes (80ul total) *Nanodrop: 32.2 ng/ul -> 32.2ng/ul / (228bp*660Da/bp) = 214nM *Dilute 2.34ul in 47.66ul H2O *Final volumes: **75ul of 214nM **50ul of 10nM
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