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#!/usr/bin/perl
use warnings;
...
  "Seg sites\tSingletons\tPi\tTheta\tTajima's D\tFu & Li F*\tFu & Li D
+*";
say $out_file
  "$segsites\t$singletons\t$pi\t$theta\t$tajima_D\t$F_star\t$D_star";

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Can't call method "isa" on unblessed reference at /usr/local/share/per
+l/5.14.2/Bio/PopGen/Statistics.pm line 901, <GEN0> line 3