nanoporetech/katuali

mini_assemble (racon) returning empty polished sequence

Linkrw12 opened this issue · 0 comments

I'm writing a custom pipeline that closely resembles all_fast_assm_polish. I have a proper assembly using mini_assemble for de novo assembly from my guppy generated basecalls. However, when I try to polish the assembly, it returns an empty polish sequence. Code below:

# -m 2 for simplicity as anything greater than 2 produces similar errors
mini_assemble -i guppy_basecalls.fastq -r consensus_draft.fasta -o racon -t 8 -p assm

Gives me the following error:

Copying FASTX input to workspace: test/guppy/test.fastq > racon/assm.fa.gz
Skipped adapter trimming.
Skipped pre-assembly correction.
Using supplied reference to perform reference-guided consensus.
Warning: Parts of this reference might not be polished by racon and such parts will be present in the final consensus unmodified.
Running racon read shuffle 1...
Running round 1 consensus...
[M::mm_idx_gen::0.003*5.27] collected minimizers
[M::mm_idx_gen::0.004*5.48] sorted minimizers
[M::main::0.004*5.46] loaded/built the index for 1 target sequence(s)
[M::mm_mapopt_update::0.005*5.01] mid_occ = 7
[M::mm_idx_stat] kmer size: 15; skip: 10; is_hpc: 0; #seq: 1
[M::mm_idx_stat::0.005*4.73] distinct minimizers: 11031 (99.27% are singletons); average occurrences: 1.010; average spacing: 5.365
[M::worker_pipeline::0.038*4.41] mapped 162 sequences
[M::main] Version: 2.17-r941
[M::main] CMD: minimap2 -K 500M -t 8 /home/robertlink/Non-thesis-work/HIV_nanopore_pipeline/test/guppy/assembly/test.fa assm.fa.gz
[M::main] Real time: 0.039 sec; CPU: 0.170 sec; Peak RSS: 0.010 GB
[racon::Polisher::initialize] loaded target sequences 0.000369 s
[racon::Polisher::initialize] loaded sequences 0.016006 s
[racon::Polisher::initialize] loaded overlaps 0.000370 s
[racon::Polisher::initialize] aligning overlaps [====================] 0.181798 s
[racon::Polisher::initialize] transformed data into windows 0.000289 s
Running round 2 consensus...
[M::mm_idx_gen::0.000*5.18] collected minimizers
[M::mm_idx_gen::0.001*4.71] sorted minimizers
[M::main::0.001*4.66] loaded/built the index for 0 target sequence(s)
[M::mm_mapopt_update::0.001*4.52] mid_occ = 1
[M::mm_idx_stat] kmer size: 15; skip: 10; is_hpc: 0; #seq: 0
[M::mm_idx_stat::0.001*4.35] distinct minimizers: 0 (-nan% are singletons); average occurrences: -nan; average spacing: -nan
[M::worker_pipeline::0.021*2.88] mapped 162 sequences
[M::main] Version: 2.17-r941
[M::main] CMD: minimap2 -K 500M -t 8 racon_1_1.fa.gz assm.fa.gz
[M::main] Real time: 0.021 sec; CPU: 0.060 sec; Peak RSS: 0.007 GB
[racon::Polisher::initialize] error: empty target sequences set!
Waiting for cleanup.
Final assembly written to racon/assm_final.fa. Have a nice day.

Even when I use -m 1 it still returns a blank output file. Is there something that I'm misunderstanding? What are your thoughts on the manner?