Hello,
It is my first time using Picard to remove duplicates, and here are some of the duplication metrics:
ESTIMATED_LIBRARY_SIZE: 24195090
PERCENT_DUPLICATION: 0.707214 (so ~70%)
READ_PAIR_DUPLICATES: 55689797 (~55.7M)
Histogram BIN1: 8692285 (so this is essentially the fragments present a single time)
In the metrics or the output file, how can I find the number of reads/fragments that have been removed?
Any help would be appreciated.
1 answer
From the metrics, the number of duplicates which were detected is the 55M number (READ_PAIR_DUPLICATES). You can find more information on the picard outputs here: https://broadinstitute.github.io/picard/picard-metric-definitions.html#DuplicationMetrics. If run in default mode the program won't automatically remove duplicates, so you would need to add REMOVE_DUPLICATES=true.
Particularly for RNA-seq, though, whether to remove PCR duplicates at all is still debated and can sometimes do more harm than good: https://www.nature.com/articles/srep25533.
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