Tag Archives: Panopea generosa

Kmer Estimation – Kmergenie on Geoduck Sequence Data (default settings)

After the last SparseAssembler assembly completed, I wanted to do another run with a different kmer size (last time was arbitrarily set at 101). However, I didn’t really know how to decide, particularly since this assembly consisted of mixed read lenghts (50bp and 100bp). So, I ran kmergenie on all of our geoduck (Panopea generosa) sequencing data in hopes of getting a kmer determination to apply to my next assembly.

The job was run on our Mox HPC node.

Slurm script: 20180419_kmergenie_geoduck_slurm.sh

Input files list (needed for kmergenie command – see Slurm script linked above): geoduck_fastq_list.txt

Results:

Output folder: 20180419_kmergenie_geoduck/

Slurm output file: 20180419_kmergenie_geoduck/slurm-161551.out

Kmer histograms (HTML): 20180419_kmergenie_geoduck/histograms_report.html

Screen cap from Kmer report:

This data estimates the best kmer size for this data to be 121.

However, based on the kmergenie documentation, this is likely to be inaccurate. This inaccuracy is based on the fact that our kmer graph should be concave. Our graph, instead, is only partial – we haven’t reached a kmer size where the number of kmers is decreasing.

As such, I’ll try re-running with a different maximum kmer settting (default max is 121).

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Assembly Stats – Quast Stats for Geoduck SparseAssembler Job from 20180405

The geoduck genome assembly started 20180405 completed this weekend.

This assembly utilized the BGI data and all of the Illumina project data (NMP and NovaSeq) with a kmer 101 setting.

I ran Quast to gather some assembly stats, using the following command:

python /home/sam/software/quast-4.5/quast.py -t 24 /mnt/owl/Athaliana/20180405_sparseassembler_kmer101_geoduck/Contigs.txt
Results:

Quast output folder: results_2018_04_15_13_45_03/

Quast report (HTML): results_2018_04_15_13_45_03/report.html

I’ve embedded the Quast HTML report below, but it may be easier to view by using the link above.

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Genome Assembly – SparseAssembler Geoduck Genomic Data, kmer=101

UPDATE 20180413

Assembly complete. See end of post for data locations.


UPDATE 20180410

Received a status update email:

SLURM Job_id=156637 Name=20180405_sparse_assembler_kmer101_geo Ended, Run time 4-20:17:08, CANCELLED, ExitCode 0

After talking to Steven, it turns out Mox was taken offline for maintenance, which killed all jobs (and access). Ugh.

Will restart tonight once Mox is back online.


OK, here we go! Initiatied an assembly run using SparseAssembler on our Mox HPC node on all of our geoduck genomic sequencing data:

Kmer size set to 101.

This is 118 files of sequencing data!! Fingers crossed…

Slurm script: 20180405_sparse_assembler_kmer101_geoduck_slurm.sh


#!/bin/bash
## Job Name
#SBATCH --job-name=20180405_sparse_assembler_kmer101_geo
## Allocation Definition
#SBATCH --account=srlab
#SBATCH --partition=srlab
## Resources
## Nodes (We only get 1, so this is fixed)
#SBATCH --nodes=1
## Walltime (days-hours:minutes:seconds format)
#SBATCH --time=30-00:00:00
## Memory per node
#SBATCH --mem=500G
##turn on e-mail notification
#SBATCH --mail-type=ALL
#SBATCH --mail-user=samwhite@uw.edu
## Specify the working directory for this job
#SBATCH --workdir=/gscratch/scrubbed/samwhite/20180405_sparseassembler_kmer101_geoduck

/gscratch/srlab/programs/SparseAssembler/SparseAssembler 
LD 0 
NodeCovTh 1 
EdgeCovTh 0 
k 101 
g 15 
PathCovTh 100 
GS 2200000000 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/AD002_S9_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/AD002_S9_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/AD002_S9_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/AD002_S9_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR005_S4_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR005_S4_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR005_S4_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR005_S4_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR006_S3_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR006_S3_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR006_S3_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR006_S3_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR012_S1_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR012_S1_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR012_S1_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR012_S1_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR013_AD013_S2_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR013_AD013_S2_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR013_AD013_S2_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR013_AD013_S2_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR014_AD014_S5_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR014_AD014_S5_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR014_AD014_S5_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR014_AD014_S5_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR015_AD015_S6_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR015_AD015_S6_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR015_AD015_S6_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR015_AD015_S6_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR019_S7_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR019_S7_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR019_S7_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR019_S7_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR021_S8_L001_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR021_S8_L001_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR021_S8_L002_R1_001_val_1_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/20180129_trimgalore_geoduck_novaseq/NR021_S8_L002_R2_001_val_2_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/151114_I191_FCH3Y35BCXX_L1_wHAIPI023989-79_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/151114_I191_FCH3Y35BCXX_L1_wHAIPI023989-79_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/151114_I191_FCH3Y35BCXX_L2_wHAMPI023988-81_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/151114_I191_FCH3Y35BCXX_L2_wHAMPI023988-81_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/151122_I136_FCH3L2FBBXX_L7_wHAXPI023990-97_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/151122_I136_FCH3L2FBBXX_L7_wHAXPI023990-97_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L3_WHPANwalDDAADWAAPEI-101_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L3_WHPANwalDDAADWAAPEI-101_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L4_WHPANwalDDAADWAAPEI-101_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L4_WHPANwalDDAADWAAPEI-101_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L5_WHPANwalDDABDLAAPEI-100_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L5_WHPANwalDDABDLAAPEI-100_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L5_WHPANwalDDACDTAAPEI-102_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L5_WHPANwalDDACDTAAPEI-102_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L6_WHPANwalDDABDLAAPEI-100_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L6_WHPANwalDDABDLAAPEI-100_2.fastq 
i1 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L6_WHPANwalDDACDTAAPEI-102_1.fastq 
i2 /gscratch/scrubbed/samwhite/bgi_geoduck/160103_I137_FCH3V5YBBXX_L6_WHPANwalDDACDTAAPEI-102_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-1_S1_L001_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-1_S1_L001_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2_S5_L002_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2_S5_L002_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-1_S2_L001_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-1_S2_L001_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-2_S6_L002_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-2_S6_L002_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-3_S10_L003_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-3_S10_L003_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-4_S14_L004_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-4_S14_L004_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-5_S18_L005_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-5_S18_L005_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-6_S22_L006_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-6_S22_L006_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-7_S26_L007_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-7_S26_L007_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-8_S30_L008_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-2to4kb-8_S30_L008_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-3_S9_L003_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-3_S9_L003_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-4_S13_L004_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-4_S13_L004_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5_S17_L005_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5_S17_L005_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-1_S3_L001_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-1_S3_L001_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-2_S7_L002_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-2_S7_L002_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-3_S11_L003_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-3_S11_L003_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-4_S15_L004_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-4_S15_L004_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-5_S19_L005_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-5_S19_L005_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-6_S23_L006_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-6_S23_L006_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-7_S27_L007_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-7_S27_L007_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-8_S31_L008_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-5to7kb-8_S31_L008_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-6_S21_L006_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-6_S21_L006_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-7_S25_L007_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-7_S25_L007_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8_S29_L008_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8_S29_L008_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-1_S4_L001_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-1_S4_L001_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-2_S8_L002_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-2_S8_L002_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-3_S12_L003_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-3_S12_L003_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-4_S16_L004_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-4_S16_L004_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-5_S20_L005_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-5_S20_L005_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-6_S24_L006_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-6_S24_L006_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-7_S28_L007_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-7_S28_L007_R2_001_val_2.fastq 
i1 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-8_S32_L008_R1_001_val_1.fastq 
i2 /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/Geoduck-NMP-gDNA-8to10kb-8_S32_L008_R2_001_val_2.fastq
Results:

Output folder: 20180405_sparseassembler_kmer101_geoduck/

Slurm output files:

SparseAssembler Assembly (FASTA): Contigs.txt

Added this to the GitHub wiki for our geoduck genome assemblies.

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Gunzip – BGI HiSeq Geoduck Genome Sequencing Data

In preparation to run SpareAssembler, I needed to gunzip the BGI gzipped FASTQ files from 20180327.

Ran the following slurm script on our Mox node:


#!/bin/bash
## Job Name
#SBATCH --job-name=20180405_geoduck_bgi_gunzip
## Allocation Definition
#SBATCH --account=srlab
#SBATCH --partition=srlab
## Resources
## Nodes (We only get 1, so this is fixed)
#SBATCH --nodes=1
## Walltime (days-hours:minutes:seconds format)
#SBATCH --time=30-00:00:00
## Memory per node
#SBATCH --mem=500G
##turn on e-mail notification
#SBATCH --mail-type=ALL
#SBATCH --mail-user=samwhite@uw.edu
## Specify the working directory for this job
#SBATCH --workdir=/gscratch/scrubbed/samwhite/bgi_geoduck

for i in /gscratch/scrubbed/samwhite/bgi_geoduck/*.gz; do
    filename="${i##*/}"
    no_ext="${filename%%.*}"
    gunzip < "$i" > "$no_ext".fastq
done
Results:

Completed in ~45mins. Will proceed with massive geoduck genome assembly!

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Gunzip – Trimmed Illumina Geoduck HiSeq Genome Sequencing Data

In preparation to run SpareAssembler, I needed to gunzip the trimmed gzipped FASTQ files from 20140401.

Ran the following slurm script on our Mox node:


#!/bin/bash
## Job Name
#SBATCH --job-name=20180404_geoduck_gunzip
## Allocation Definition
#SBATCH --account=srlab
#SBATCH --partition=srlab
## Resources
## Nodes (We only get 1, so this is fixed)
#SBATCH --nodes=1
## Walltime (days-hours:minutes:seconds format)
#SBATCH --time=30-00:00:00
## Memory per node
#SBATCH --mem=500G
##turn on e-mail notification
#SBATCH --mail-type=ALL
#SBATCH --mail-user=samwhite@uw.edu
## Specify the working directory for this job
#SBATCH --workdir=/gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck

for i in /gscratch/scrubbed/samwhite/illumina_geoduck_hiseq/20180328_trim_galore_illumina_hiseq_geoduck/*.gz; do
    filename="${i##*/}"
    no_ext="${filename%%.*}"
    gunzip < "$i" > "$no_ext".fastq
done
Results:

This crashed shortly after initiating the run (~30mins later). Received following email notification:

SLURM Job_id=155940 Name=20180404_geoduck_gunzip Failed, Run time 00:30:40, NODE_FAIL

It did not generate a slurm output file, nor any gunzipped files. Will contact UW IT…

UPDATE 20140404

Weird, about an hour after this crashed, I received the following email, indicating the job was submitted (I did no resubmit, btw):

SLURM Job_id=155940 Name=20180404_geoduck_gunzip Began, Queued time 00:02:29

Completed about 3hrs later.

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DNA Isolation & Quantification – Geoduck larvae metagenome filter rinses

This is another attempt to isolate DNA from two more of the geoduck hatchery metagenome samples Emma delivered on 20180313.

The previous attempt, using DNAzol, did not yield any DNA.

I isolated DNA from the following two samples:

  • MG 5/19 #4
  • MG 5/26 #4

I used the DNA Stool Kit (Qiagen), following the “Stool Human DNA” protocol with the following changes:

  • Incubated @ 95oC for 5mins after initial addition of Buffer ASL. This is a lysis step that might help increase yields (see the “Stool Pathogen Detection” protocol)
  • Did not add InhibitEX Tablet. Deemed unnecessary, since these weren’t stool samples.
  • Eluted in 50μL of Buffer AE

I opted to follow the “Stool Human DNA” protocol, as it processes a larger portion of the initial sample, compared to the “Stool Pathogen Detection” protocol (600μL vs. 200μl)

Samples were quantified using the Roberts Lab Qubit 3.0 with the Qubit High Sensitivity dsDNA Kit (Invitrogen).

10μL of each sample were used.

Results:

Neither sample yielded any detectable DNA. Will discuss with Steven.

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TrimGalore!/FastQC/MultiQC – Illumina HiSeq Genome Sequencing Data Continued

The previous attempt at this was interrupted by a random glitch with our Mox HPC node.

I removed the last files processed by TrimGalore!, just in case they were incomplete. I updated the slurm script to process only the remaining files that had not been processed when the Mox glitch happened (including the files I deemed “incomplete”).

As in the initial run, I kept the option in TrimGalore! to automatically run FastQC on the trimmed output files.

TrimGalore! slurm script: 20180401_trim_galore_illumina_geoduck_hiseq_slurm.sh

MultiQC was run locally once the files were copied to Owl.

Results:

Job completed on 20180404.

Trimmed FASTQs: 20180328_trim_galore_illumina_hiseq_geoduck/

MD5 checksums: 20180328_trim_galore_illumina_hiseq_geoduck/checksums.md5

  • MD5 checksums were generated on Mox node and verified after copying to Owl.

Slurm output file: 20180401_trim_galore_illumina_geoduck_hiseq_slurm.sh

TrimGalore! output: 20180328_trim_galore_illumina_hiseq_geoduck/20180404_trimgalore_reports/

FastQC output: 20180328_trim_galore_illumina_hiseq_geoduck/20180328_fastqc_trimmed_hiseq_geoduck/

MultiQC output: 20180328_trim_galore_illumina_hiseq_geoduck/20180328_fastqc_trimmed_hiseq_geoduck/multiqc_data/

MultiQC HTML report: 20180328_trim_galore_illumina_hiseq_geoduck/20180328_fastqc_trimmed_hiseq_geoduck/multiqc_data/multiqc_report.html

Trimming completed and the FastQC results look much better than before.

Will proceed with full-blown assembly!

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FastQC/MultiQC – Illumina HiSeq Genome Sequencing Data

Since running SparseAssembler seems to be working and actually able to produce assemblies, I’ve decided I’ll try to beef up the geoduck genome assembly with the rest of our existing genomic sequencing data.

Yesterday, I transferred our BGI geoduck data to our Mox node and ran it through FASTQC

I transferred our Illumina HiSeq data sets (*NMP*.fastq.gz) to our Mox node (/gscratch/scrubbed/samwhite/illumina_geoduck_hiseq). These were part of the Illumina-sponsored sequencing project.

I verified the MD5 checksums (not documented) and then ran FASTQC, followed by MultiQC.

FastQC slurm script: 20180328_fastqc_illumina_geoduck_hiseq_slurm.sh

This was followed with MultiQC (locally, after copying the the FastQC output to Owl).

Results:

FASTQC output: 20180328_illumina_hiseq_geoduck_fastqc

MultiQC output: 20180328_illumina_hiseq_geoduck_fastqc/multiqc_data

MultiQC HTML report: 20180328_illumina_hiseq_geoduck_fastqc/multiqc_data/multiqc_report.html

Well, lots of fails. I high level of “Per Base N Content” (these are only warnings, but we also haven’t received data with these warnings before). Also, they all fail in the “Overrepresented sequences” analysis.

I’ll run these through TrimGalore! (probably twice), and see how things change.

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TrimGalore!/FastQC/MultiQC – Illumina HiSeq Genome Sequencing Data

Previous FastQC/MultiQC analysis of the geoduck Illumina HiSeq data (NMP.fastq.gz files) revealed a high level of overrepresented sequences, high levels of Per Base N Content, failure of Per Sequence GC Content, and a few other bad things.

Ran these through TrimGalore! on our Mox HPC node.

Added an option in TrimGalore! to automatically run FastQC on the trimmed output files.

TrimGalore! slurm script: 20180328_trim_galore_illumina_geoduck_hiseq_slurm.sh

Results:

Slurm output file: slurm-153098.out

I received a job status email on 20180330:

SLURM Job_id=153098 Name=20180328_trim_galore_geoduck_hiseq Failed, Run time 1-17:22:47, FAILED, ExitCode 141

The slurm output file didn’t indicate any errors, so I restarted the job and contacted UW IT to see if I could get more info.

UPDATE

Here’s their response:

04/02/2018 9:13 AM PDT – Matt

Hi Sam,

Your job died because of a networking hiccup that caused GPFS (/gscratch filesystem and such) to expel the node from the GPFS cluster. It’s a symptom of a known ongoing network issue that we’re actively working with Lenovo/Intel/IBM. Things like this aren’t happening super frequently, but enough that we recognized something was wrong and started investigating with vendors. Unfortunately, your job was unlucky and got bitten by it.

So, in short, you or your job didn’t do anything wrong. If you haven’t already (and if it is possible for your use case), we would highly recommend building in some sort of periodic state-preserving behavior (and a method to “resume”) into your longer-running jobs. Jobs can unexpectedly die for any number of reasons, and it is nice not to lose days of compute progress when that happens.

-Matt

Well, okay then.

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FastQC/MultiQC – BGI Geoduck Genome Sequencing Data

Since running SparseAssembler seems to be working and actually able to produce assemblies, I’ve decided I’ll try to beef up the geoduck genome assembly with the rest of our existing genomic sequencing data.

I transferred our BGI geoduck FASTQ files to our Mox node (/gscratch/scrubbed/samwhite/bgi_geoduck/).

I ran FASTQC on them to actually check them out and see if they needed any trimming, as I don’t believe this has been done!

FASTQC slurm script: 20180327_fastqc_bgi_geoduck_slurm.sh

Side note: Initial FASTQC failed on one file. Turns out, it got corrupted during transfer! Serves as good reminder about the importance of verifying MD5 checksums after file transfer, prior to attempting to work with files!

This was followed up with MultiQC (run locally from my computer on the files hosted on Owl). This was performed the following day (20180328).

Results:

FASTQC output: 20180327_bgi_fastqc

MultiQC output: 20180328_bgi_multiqc

MultiQC HTML report: 20180328_bgi_multiqc/multiqc_report.html

Everything looks nice and clean! Waiting on transfer and FASTQC of Illumina NMP data before proceeding to next assembly attempt.

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