Wiki » History » Revision 6
Revision 5 (Katie Lennard, 09/20/2022 11:01 AM) → Revision 6/26 (Katie Lennard, 09/20/2022 01:52 PM)
# Wiki
# Data location:
The data was transferred from Athena medmicro):
```
/MedMicro/Clinton/CRE Pfizer Feb 2022/CRE study_1A_results_17022022
/MedMicro/Clinton/CRE Pfizer Feb 2022/CRE study_1B_results_21022022
```
to Ilifu:
```
/scratch3/users/katiel/Clinton/CRE_study_August_2022/
```
# Reference data:
Klebsiella pneumoniae – strain HS11286 (GenBank accession no. CP003200.1) (n=18);
Serratia marcescens – strain KS10 (GenBank accession no. CP027798.1) (n=3);
Escherichia coli – strain ATCC 25922 (GenBank accession no. CP009072.1) (n=1); and
Enterobacter cloacae – strain ATCC 13047 (GenBank accession no. NC_014121.1) (n=1).
```
/scratch3/users/katiel/Clinton/CRE_study_August_2022/ref_genomes
```
# Objectives workflow:
![workflow.png]()
# QC:
11 sample failed QC phred scores before trimming and filtering; none failed after filtering and trimming. Filtering and trimming were executed as follows:
```
nextflow run kviljoen/fastq_QC --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/raw/study_1A_B_combined/*_R{1,2}_001.fastq.gz' -profile ilifu
```
QC reports can be found in the 'files' tab
# AMR profiling
The preference from Clinton is to do AMR profiling with the ResFinder DB. I'm getting errors there that I think relate to the header formatting though so in the interim have run with the ARG_annot DB that we used for previous projects as:
## ARGannot
```
nextflow run kviljoen/uct-srst2 --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/2022-09-19-fastq_QC/bbduk/*_{1,2}.fq' -profile ilifu --gene_db /cbio/users/katie/Nicol/Ps_aerug_srst2_MLST/ARGannot_r3.fasta --outdir /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_ARGannot/coverage_80_run /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_resFinder/coverage_80_run --min_gene_cov 80
```
## CARD DB:
This database is the recommended by srst2 and has been formatted by them already. The DB was downloaded with:
```
wget https://github.com/katholt/srst2/blob/master/data/CARD_v3.0.8_SRST2.fasta?raw=true -O CARD_v3.0.8_SRST2.fasta
```
Pipeline execution as:
```
nextflow run kviljoen/uct-srst2 --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/2022-09-19-fastq_QC/bbduk/*_{1,2}.fq' -profile ilifu --gene_db /scratch3/users/katiel/Clinton/CRE_study_August_2022/ref_files/CARD_v3.0.8_SRST2.fasta --outdir /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_CARD/coverage_80_run --min_gene_cov 80
```
# Data location:
The data was transferred from Athena medmicro):
```
/MedMicro/Clinton/CRE Pfizer Feb 2022/CRE study_1A_results_17022022
/MedMicro/Clinton/CRE Pfizer Feb 2022/CRE study_1B_results_21022022
```
to Ilifu:
```
/scratch3/users/katiel/Clinton/CRE_study_August_2022/
```
# Reference data:
Klebsiella pneumoniae – strain HS11286 (GenBank accession no. CP003200.1) (n=18);
Serratia marcescens – strain KS10 (GenBank accession no. CP027798.1) (n=3);
Escherichia coli – strain ATCC 25922 (GenBank accession no. CP009072.1) (n=1); and
Enterobacter cloacae – strain ATCC 13047 (GenBank accession no. NC_014121.1) (n=1).
```
/scratch3/users/katiel/Clinton/CRE_study_August_2022/ref_genomes
```
# Objectives workflow:
![workflow.png]()
# QC:
11 sample failed QC phred scores before trimming and filtering; none failed after filtering and trimming. Filtering and trimming were executed as follows:
```
nextflow run kviljoen/fastq_QC --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/raw/study_1A_B_combined/*_R{1,2}_001.fastq.gz' -profile ilifu
```
QC reports can be found in the 'files' tab
# AMR profiling
The preference from Clinton is to do AMR profiling with the ResFinder DB. I'm getting errors there that I think relate to the header formatting though so in the interim have run with the ARG_annot DB that we used for previous projects as:
## ARGannot
```
nextflow run kviljoen/uct-srst2 --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/2022-09-19-fastq_QC/bbduk/*_{1,2}.fq' -profile ilifu --gene_db /cbio/users/katie/Nicol/Ps_aerug_srst2_MLST/ARGannot_r3.fasta --outdir /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_ARGannot/coverage_80_run /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_resFinder/coverage_80_run --min_gene_cov 80
```
## CARD DB:
This database is the recommended by srst2 and has been formatted by them already. The DB was downloaded with:
```
wget https://github.com/katholt/srst2/blob/master/data/CARD_v3.0.8_SRST2.fasta?raw=true -O CARD_v3.0.8_SRST2.fasta
```
Pipeline execution as:
```
nextflow run kviljoen/uct-srst2 --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/2022-09-19-fastq_QC/bbduk/*_{1,2}.fq' -profile ilifu --gene_db /scratch3/users/katiel/Clinton/CRE_study_August_2022/ref_files/CARD_v3.0.8_SRST2.fasta --outdir /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_CARD/coverage_80_run --min_gene_cov 80
```