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We used hard filter for removing variants having below 5X coverage to reduce false positives.
Plus sign and down triangle are additionally filtered by the GATK hard filter.
Second, a physicochemical properties hard filter was applied to compounds with calculated log P>6 and Mr>600 Da.
Variants were predicted using GATK [ 6] (V2.4) using default parameters and filtered using the GATK hard filter.
Using the GATK hard filter, only 28%% of the 353,076 filtered SNVs are concordant between capture kits.
It is often unclear what an adequate hard filter is; beyond general guidelines each specific study requires experimenting and empirical testing.
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SNPs were filtered using VQSR and indels were hard filtered using GATK version 2.2 variant filtration.
Hard filters were set according to the GATK Best Practices recommendations [ 32] for GATK called variant sets.
A possible solution for selecting the appropriate list of variants is the use of hard filters.
Hard filtering was applied to indels and only passing indels were used for subsequent analyses.
The non-uniform coverage often seen in NGS studies [ 8] makes hard filtering a challenging task.
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