nanoporetech / pipeline-umi-amplicon

Workflow to prepare high accuracy single molecule consensus sequences from amplicon data using unique molecular identifiers
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all reads are short? #36

Open maxstengl opened 2 months ago

maxstengl commented 2 months ago

Hello, on my test data the pipeline doesn't run past alignment ERROR - did not find a single cluster passing the min_read threashold! I assume this is because of all candidate reads are considered short as the umi stats file is written as:

The reads are 100-300 bases long the region in the bed file is chr12 25398185 25398385 KRAS_Target min_overlap 0.01 and 0.1 did not work all resulted in:

Reads found: 41943 Reads unmapped: 1959 (4%) KRAS_Target Reads found: 0 On target: 66 (0%) 0 concatamers - 0% 66 short - 100%

I changed the min_overlap parameter in the yml file and even tried -1 and 0 but nothing changes the outcome. We do not use ont UMIs but I adapted the yml file accordingly - but maybe this causes the root problem as no reads to cluster are found? I guess I am missing something simple - please help ! Thank you, Daniel