Comments (6)
from gembs.
Hello Simon,
We have a library which was prepared for 75PE sequencing so the insert size was short. We also sequenced this library with 150PE on NovaSeq to test the instrument. As expected, there is a lot of adapter contamination in the 150PE data. Although gem3-mapper aligns most of the reads regardless of the adapter contamination, the rate of unique reads decreases considerably. For the same sample the average unique rate increases from 44% to 54% after adapter+3'quality trimming whereas it is 74% in the 75PE data. The rate for 75PE is much higher because GEM3 mapper gives 0 mapping quality to 100% overlapping pairs. Moreover, the adapters are aligned with gaps and mismatches rather than soft clipping which might -IMHO- increase the FDR of variant/bisulfite calling. Please have a look at the IGV images below where you can see the difference between 75PE, adapter trimmed 150PE and non-trimmed 150PE data (from top to bottom respectively).
https://drive.google.com/file/d/1oG7Y1wRxKEjarDN5HC1qg_MkuIxpqztQ/view?usp=sharing
https://drive.google.com/file/d/1yrXTMHkK1xZRz1BWcBe03Qit7BeBr-Fa/view?usp=sharing
Best,
Bekir
from gembs.
from gembs.
Thank you Simon, it would be great if you can implement this soon.
Best, Bekir
from gembs.
Hello Simon,
I could add a flag to the caller to not discard such read pairs (100% overlapping)
Did you have a chance to implement this feature?
Best, Bekir
from gembs.
Hi Bekir,
In principle the new version of gemBS (3.0.0) should have this feature. The --keep-unmatched option to gemBS call will not discard read pairs that are not pairing correctly.
Simon
from gembs.
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from gembs.