BioMicroCenter:PPR Program: Difference between revisions

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<H2> PPR PROGRAM </H2>
<H2> PPR PROGRAM </H2>
Guaranteeing high quality next-generation sequencing (NGS) data in a rapidly changing environment is an ongoing challenge. The recent introduction of the [BioMicroCenter:Sequencing|Illumina NextSeq500] and the depreciation of specific metrics from Illumina's Sequencing Analysis Viewer (SAV) have made it more difficult to directly determine the baseline error rate of sequencing runs. We have created an open-source tool to construct the Percent Perfect Reads (PPR) plot previously provided by the Illumina sequencers. The PPR program is compatible with HiSeq2000/2500, MiSeq, and NextSeq500 instruments, and provides an alternative to Illumina's Q scores for determining run quality.  
Guaranteeing high quality next-generation sequencing (NGS) data in a rapidly changing environment is an ongoing challenge. The recent introduction of the [[BioMicroCenter:Sequencing|Illumina NextSeq500]] and the depreciation of specific metrics from Illumina's Sequencing Analysis Viewer (SAV) have made it more difficult to directly determine the baseline error rate of sequencing runs. We have created an open-source tool to construct the Percent Perfect Reads (PPR) plot previously provided by the Illumina sequencers. The PPR program is compatible with HiSeq2000/2500, MiSeq, and NextSeq500 instruments, and provides an alternative to Illumina's Q scores for determining run quality.  


<LI> PPR Program can be downloaded as tarball (.tgz) file [[Media:NGS_sequencing_QC.zip|'''here''']]<br>
<LI> PPR Program can be downloaded as tarball (.tgz) file [[Media:NGS_sequencing_QC.zip|'''here''']]<br><BR>


The software is designed to be run in a UNIX/LINUX environment.  
The software is designed to be run in a UNIX/LINUX environment.  
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<H3>GENERIC:</H3>
<H3>GENERIC:</H3>
  cd [CODE DIRECTORY]
   perl ./NGS_missmatch_qc.pl [RUNTYPE] (FASTQ FILES)
   perl ./NGS_missmatch_qc.pl [RUNTYPE] (FASTQ FILES)



Revision as of 07:27, 17 May 2016

HOME -- SEQUENCING -- LIBRARY PREP -- HIGH-THROUGHPUT -- COMPUTING -- OTHER TECHNOLOGY

PPR PROGRAM

Guaranteeing high quality next-generation sequencing (NGS) data in a rapidly changing environment is an ongoing challenge. The recent introduction of the Illumina NextSeq500 and the depreciation of specific metrics from Illumina's Sequencing Analysis Viewer (SAV) have made it more difficult to directly determine the baseline error rate of sequencing runs. We have created an open-source tool to construct the Percent Perfect Reads (PPR) plot previously provided by the Illumina sequencers. The PPR program is compatible with HiSeq2000/2500, MiSeq, and NextSeq500 instruments, and provides an alternative to Illumina's Q scores for determining run quality.

  • PPR Program can be downloaded as tarball (.tgz) file here

    The software is designed to be run in a UNIX/LINUX environment.

    Dependencies:

  • fastxtoolkit (confirmed for v0.0.13)
  • bowtie2 (confirmed for v2.2.3)
  • bedtools (confirmed for v2.20.1)
  • r (confirmed for v2.15.3)
  • perl (confirmed for v5.20.0)

    Commands:

    GENERIC:

     cd [CODE DIRECTORY]
     perl ./NGS_missmatch_qc.pl [RUNTYPE] (FASTQ FILES)
    

    SPECIFIC:

    For Paired end NextSeq sequencing:

     perl ./NGS_missmatch_qc.pl nextseq_paired_end read1.fq read2.fq
    

    For single end NextSeq sequencing:

     perl ./NGS_missmatch_qc.pl nextseq_single_end read1.fq
    

    For paired end MiSeq sequencing:

     perl ./NGS_missmatch_qc.pl miseq_paired_end read1.fq read2.fq
    

    For single end MiSeq sequencing

     perl ./NGS_missmatch_qc.pl miseq_single_end read1.fq
    

    For paired end HiSeq sequencing

     perl ./NGS_missmatch_qc.pl hiseq_paired_end Lane1_1.fq Lane1_2.fq Lane2_1.fq Lane2_2.fq Lane3_1.fq Lane3_2.fq 
     Lane4_1.fq Lane4_2.fq Lane5_1.fq Lane5_2.fq Lane6_1.fq Lane6_2.fq Lane7_1.fq Lane7_2.fq Lane8_1.fq Lane8_2.fq 
    

    For single end HiSeq sequencing

     perl ./NGS_missmatch_qc.pl hiseq_single_end Lane1.fq Lane2.fq Lane3.fq Lane4.fq Lane5.fq Lane6.fq Lane7.fq Lane8.fq