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===Day 3=== #Wash with nfH2O twice #Prepare Ampligase mix on ice #*Preheat H2O + Padlock probes + suppv2 Oligos to 85C and then snap cool before adding Ampligase buffer and enzyme *B13: 305nM **This batch of padlock probes was PCR'd an extra 3 cycles during production PCR {| {{table}} | align="center" style="background:#f0f0f0;"|'''Component''' | align="center" style="background:#f0f0f0;"|'''Volume''' |- | DEPC-H2O||18.43 |- | Ampligase Buffer||10 |- | Erin's Batch 13 2/12/2016 Padlock Probes 305nM]]||33 |- | 100nM suppv2 Oligos || 28.57 |- | Ampligase||10 |- | Total||100 |} #Add mix to sample and incubate for 30min at 37C #Move sample to 60C and oven slowly decreases to 55C and held for ~29hr
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