1. Fully mask genome a. customize a repeat library if do not have one $ BuildDatabase -name dbname -engine wublast refSeq.fasta $ RepeatModeler -pa 32 -engine wublast -database dbname b. Identify segmental duplications in reference genome $ WGAC_SD.pl -i refSeq.fasta -l repeat.lib.fasta -t 48 c. thoroughly mask reference genome and get a unique genome $ thorough_mask.pl -i ref.fasta -r TEseq.fasta -t 48 -s SD_interval.bed 2. Mapping reads to unique genome mrfast 3. calculate and normalize read depth for each sample 4. call CNVs and calulate gene copy number 5. compare allele frequency