Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • epistatic
    Senior Member
    • Mar 2009
    • 129

    Information provided to users by sequencing sites

    Hello all,

    Data generation at our site is now a non-issue and I see a huge need to standardize the QC of sequencing data before handing off to customers. We have several different options for baseline QC and specialized QC information based upon library source: RNA-seq, Exome-seq, ChIP-seq, etc. I also believe it is hard to know the true quality without some initial alignment. This can get computationally expensive with the sample #s we are starting to have from multiplexed runs on the HiSeqs but I still think should be done.

    Does anyone have recommendations on what information is essential? Are there any posted standards more savvy than Illumina's 80% of bases >Q30?
  • NextGenSeq
    Senior Member
    • Apr 2009
    • 482

    #2
    The HiSeq aligns the PhiX control to PhiX. If you spike in 1 to 2% that's enough to know if the flowcell or sequencing reagents were good.

    If those look good and the library doesn't it's the library (which is almost always the case) or the library was misquantified.

    Comment

    • epistatic
      Senior Member
      • Mar 2009
      • 129

      #3
      Yes, PhiX tells you the instrument is working...

      I apologize if my question was vague. I am interested in knowing what information deliverables would be the most useful for users to receive with their sequencing data. Providing users with FASTQ files and the CASAVA demultiplexing summaries for the data (yield, read count, PF, %>/= Q30) seems reasonable but also not completely informative.

      Comment

      • kopi-o
        Senior Member
        • Feb 2008
        • 319

        #4
        Mapping rate to the genome, % duplicates, fraction mRNA (for RNA-seq), % reads on target (exome / seq cap) seem useful.

        Comment

        Latest Articles

        Collapse

        • GATTACAT
          Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
          by GATTACAT
          Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
          07-01-2026, 11:43 AM
        • SEQadmin2
          Nine Things a Sample Prep Scientist Thinks About Before Sequencing
          by SEQadmin2


          I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.

          Here are nine questions we think about, in roughly the order they matter, before...
          06-18-2026, 07:11 AM

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by SEQadmin2, 07-02-2026, 11:08 AM
        0 responses
        12 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-30-2026, 05:37 AM
        0 responses
        14 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-26-2026, 11:10 AM
        0 responses
        20 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-17-2026, 06:09 AM
        0 responses
        54 views
        0 reactions
        Last Post SEQadmin2  
        Working...