Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • ramirob
    Member
    • Apr 2012
    • 16

    [sam_read1] reference 'chrN.fa' is recognized as '*'

    Hello,

    I aligned some human samples with Casava, and then converted the alignments to SAM using samtools:

    llumina_export2sam.pl --read1=file_R1.export.txt.gz --read2=file_R2.export.txt.gz > file.sam

    Then I try to convert to bam:

    samtools view -bt genome.fa.fai file.sam > file.bam

    but I get a lot of

    [sam_read1] reference 'chr8.fa' is recognized as '*'.
    [sam_read1] reference 'chr11.fa' is recognized as '*'.
    etc.

    There was a similar thread that suggested changing the header as follows:

    The first line needs to be header.
    The second line needs to be a dummy read group line
    The next lines need to contain the chromosomes and their lengths.

    An example of a good header is as follows:

    @HD VN:1.0 SO:unsorted
    @RG ID:unknownReadGroup SM:unknownSample
    @SQ SN:chrI AS:ce6_32r_index LN:15072421
    @SQ SN:chrII AS:ce6_32r_index LN:15279323
    @SQ SN:chrIII AS:ce6_32r_index LN:13783681
    @SQ SN:chrIV AS:ce6_32r_index LN:17493785
    @SQ SN:chrM AS:ce6_32r_index LN:13794
    @SQ SN:chrV AS:ce6_32r_index LN:20919568
    @SQ SN:chrX AS:ce6_32r_index LN:1771885


    where do I get all of that information? I also might have to do this over and over again so I was looking for some software or precise ways so that I can write a script to do it.

    Thanks in advance,

    Ramiro
  • naman.neoanderson007
    Junior Member
    • Mar 2013
    • 1

    #2
    Hi, Ramiro

    I have just entered in 'seqanswers' and saw your question

    I suggest you to check your GENOME file - "genome.fa.fai"
    Is it in proper FASTA format ?

    Correct FASTA formated GENOME file will solve your problem.

    Comment

    Latest Articles

    Collapse

    • 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
    • SEQadmin2
      From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
      by SEQadmin2


      Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


      The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
      ...
      06-02-2026, 10:05 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by SEQadmin2, 06-26-2026, 11:10 AM
    0 responses
    12 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-17-2026, 06:09 AM
    0 responses
    48 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-09-2026, 11:58 AM
    0 responses
    107 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-05-2026, 10:09 AM
    0 responses
    125 views
    0 reactions
    Last Post SEQadmin2  
    Working...