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  • poor quality on R1 end in SOLid fastqc test

    It's my first time dealing with Solid data. It's mate pair reads, solid sequenced. A I found some problem after I used fastqc to check the raw reads, which were SRA data downloaded from NCBI:

    1. The qualities for R1 and R2 were imbalance, especially showing very poor quality in R1. What might be the problem? The only thing I did before it was converting sra format to fastq.

    2. Why are there so many Ns in R1 data? Is it because fastqc doesn't know how to treat dot in colorspace codes?

    3. Someone tried to align those reads onto ref genome. It came out very few reads could be mapped. At most 50% of the reads could be mapped. We used aligner BFAST, and also tried BWA. Since in the publication, this dataset has a good aligment, it's impossible due to bad sequencing.

    Could anyone give me some suggestion?

  • #2
    poor quality on R1 end in SOLid fastqc test

    I worked with some single end SOLiD data a couple of years ago.

    At that time the Life Tech practice was not to filter the reads (say in the way that Illumina does), and let the mapping filter out the bad reads. So as a result you got a huge number of reads, but you would expect a lower percentage of reads to align.

    The reads at the edges of the flow cell ( start and end of fastq file) had lots of Ns in them.

    Comment


    • #3
      Originally posted by mastal View Post
      I worked with some single end SOLiD data a couple of years ago.

      At that time the Life Tech practice was not to filter the reads (say in the way that Illumina does), and let the mapping filter out the bad reads. So as a result you got a huge number of reads, but you would expect a lower percentage of reads to align.

      The reads at the edges of the flow cell ( start and end of fastq file) had lots of Ns in them.
      Thanks, mastal.

      In your point, Life Tech was not to filter the "bad" reads, that caused the poor quality in the mate pairs. But why all the Ns junks are only on one end, but not the other end? This really confused me.

      Comment

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