Pooling design information should be annotated in the pif files. The pif file should have four columns (PoolBAM, Phenotype, Inds, Chroms). The PoolBAM column should specify the location of the BAM file (post pre-processing, i.e. calibration and merging) , the second column, Phenotype, should have a binary label of case/control (1/0) status or phenotypic extreme high/low (1/0) , the third column should specify the number of individuals in the corresponding pool, and the fourth column should specify the number of chromosomes in the corresponding pool.