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  • Trouble with box-plot of quality data in SOLID

    hi everyone,
    I have used the SOLiD2std.pl to change the csfasta and qual files to standard fastq files.
    I then ran fastqc to view boxplots of quality data and got these results, attached.
    They seem to have poor quality every 5 nt, as if the primer 5 of the procedure failed....
    Has anyone seen this type of quality plots before???
    I am new with solid data, have worked with ilumina and 454 before.
    Thanks for any guidance in advance.
    cheers
    maximo
    Attached Files

  • #2
    You can not convert colorspace to basespace directly. See here why:
    Discussion of next-gen sequencing related bioinformatics: resources, algorithms, open source efforts, etc

    Comment


    • #3
      An alternative route to using FastQC, which is very nice, is to align first and use the resulting BAM file as input.

      Another program I have looked at for duplicate analysis is prinseq, which also offers graphical output.

      Comment


      • #4
        Did you removed adapter info or trimmed your reads.
        what about other fastQC results they are pass or fail.
        Krishna

        Comment


        • #5
          Originally posted by Krish_143 View Post
          Did you removed adapter info or trimmed your reads.
          what about other fastQC results they are pass or fail.
          Some aspects passed and others failed,
          I did NO removal of adapters.... Not sure if it was done when i received the csfasta and qual files..
          Taking into consideration that this is a direct conversion from color to base space, its understandable that there may be confusing results.... (In all about 45% of reads mapped with color-space-novoAlign to a reference, and only 15% usnig colorspace-tophat/bowtie...)
          Any thoughts?
          PASS Basic Statistics Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          FAIL Per base sequence quality Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          WARN Per sequence quality scores Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          WARN Per base sequence content Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          FAIL Per base GC content Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          FAIL Per sequence GC content Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          WARN Per base N content Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          PASS Sequence Length Distribution Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          PASS Sequence Duplication Levels Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          PASS Overrepresented sequences Corrida_4_FC_1_01_01CVATFR001_F3.fastq
          FAIL Kmer Content Corrida_4_FC_1_01_01CVATFR001_F3.fastq

          Comment


          • #6
            I would recommend using SAET to clear up any colour errors before alignment. Normally I get 1-5% more aligned reads using this (apparently a lot more with ECC data).

            I don't think your fastqc boxplot results are too bad, but I've seen better ones.

            Comment

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