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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 can be cheaper.   ⁶Our sequencing cost estimate assumesto  purchase of an Illumina MiSeq run from a sequencing facility.