User guide

Quick start

ACEPipe is installed in ACE HPC clusters. To activate ACEPipe:

$ source activate acepipe

To get the help or available tools in ACEPipe:

$ acepipe -h
$ acepipe --help

Which will return the available commands available:

Usage: acepipe [OPTIONS] COMMAND [ARGS]...

Options:
  -h, --help  Show this message and exit.

Commands:
  makedb
  runpipe
  runqc
  runsingle

To get further options available in the command:

$ acepipe runpipe -h
$ acepipe runpipe --help

Which will return all options available for the command:

Usage: acepipe runpipe [OPTIONS]

Options:
  -h, --help                      Show this message and exit.
  -s, --sheetfile TEXT            provide sheetfile location
                                  default=samplesheet.tab
  -o, --outdir TEXT               provide output folder location
                                  default=acepipe_results
  -e, --email TEXT                provide email id
  -dt, --taxonomydb TEXT          provide reference taxonomy database location
                                  example:
                                  /usr/local/qiime_db/silva_taxonomy_db.qza
                                  default=silvadb & unitedb
  -dg, --genomedb TEXT            provide reference sequence database location
                                  example: /usr/local/qiime_db/silva_seq_db.qz
                                  default=silvadb & unitedb
  -qm, --qiimemode [forward|both]
                                  provide qiime reads mode
                                  default=forward
  -l, --readlength INTEGER        provide reads length to keep after trimming
                                  default=250
  -q, --quality INTEGER           provide quality score threshold value
                                  default=15
  -j, --joblist TEXT              provide job id as a comma-separated list
                                  default=all

Input

Input file for ACEPipe is the samplesheet file which is generated by ACE LIMS and provided by the ACE Sequencing Team. Samplesheet is a tab delimited file which contains some information such as:

  • Sample_ID
  • Sample_Name
  • Sample_Plate
  • Sample_Well
  • I7_Index_ID
  • index
  • I5_Index_ID
  • index2
  • Sample_Project
  • Description
  • Read 1 File
  • Read 2 File
  • Fwd_Primer_Name
  • Fwd_Primer_TS_seq
  • Rev_Primer_Name
  • Rev_Primer_TS_seq
  • Client_Sample_Name

ACEPipe expects the filename should shart with sample id. If your file name starts with job id, please rename the files. The fastq file name format is CasavaOneEightSingleLanePerSampleDirFmt which naming is as '.+_.+_L[0-9][0-9][0-9]_R[12]_001\.fastq\.gz'. So the name of your files should be as: SAMPLEID_JOBID_OTHERID_L001_R1_001.fastq.gz. Example: SB2658_J1235_S352_L001_R1_001.fastq.gz and SB2658_J1235_S352_L001_R2_001.fastq.gz

Output

ACEPipe wraps all the files with a html file. Transfer the result folder to your local computer and open the index.html file in any internet browser (Google Chrome preferred).

Examples

To activate ACEPipe:

$ source activate acepipe

Example-1: Forward reads processing in QIIME2

To process only the forwards reads in QIIME2:

$ nohup acepipe runpipe -s samplesheet.tab -o output_folder -e id@eaxmple.com > acepipe_forward_output.log &

Example-2: Both reads processing in QIIME2

To process both reads in QIIME2:

$ nohup acepipe runpipe -s samplesheet.tab -o output_folder -qm both -e id@eaxmple.com > acepipe_both_output.log &

Example-3: Customized database

To create customized database:

$ acepipe makedb -dg location/to/reference/sequences/file -dt location/to/taxonomy/file -o output/to/custom/database

To use the customized database:

$ nohup acepipe runpipe -s samplesheet.tab -dg location/to/reference/sequences/file -dt location/to/taxonomy/file
-o output_folder > acepipe_forward_customDB_output.log &

Example-4: Customized quality control

To process with q value of 25 and reads length of 220:

$ nohup acepipe runpipe -s samplesheet.tab -o output_folder -q 25 -l 220 -e id@eaxmple.com > acepipe_forward_customQC_output.log &

Example-5: Single or multiple jobs processing

ACEPipe processes all jobs by default. However, a single job or multipe jobs can be processed. To process a single job:

$ nohup acepipe runpipe -s samplesheet.tab -o output_folder -J JOBID -e id@eaxmple.com > acepipe_forward_JOBID_output.log &

To process multiple jobs:

$ nohup acepipe runpipe -s samplesheet.tab -o output_folder -J JOBID1,JOBID2,JOBID3,JOBIDX -e id@eaxmple.com > acepipe_forward_JOBS_output.log &

Example-6: Single reads

To use only single reads:

$ nohup acepipe runsingle -s samplesheet.tab -o output_folder -rd forward > acepipe_single_forward_output.log &

Note

Please provide your valuable comments and suggestions to make ACEPipe better. If you need further help, please contact Dr. Shihab Hasan (shihab.hasan@uq.net.au).