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²This assumes purchase of a standard 200U PCR reagent kit.   ³PCR cleanup can be performed in a number of ways; gel extraction, beads, or columns for example. Here we assume purchase of a standard column-based kit.   ⁴Sanger sequencing cost is given as the price per reaction ($7 at our sequencing facility), times the forward and reverse reactions.   ⁵This assumes the purchase of a 48-sample Nextera or TrueSeq kit from Illumina, however kits from other manufacterers manufacturers  can be cheaper. ⁶Our sequencing cost estimate assumes purchase of an Illumina MiSeq run from a sequencing facility.