Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • AdaLovelace
    Junior Member
    • Dec 2015
    • 2

    Mate pair de novo assembly

    Hi everyone, i am having some problems with a de novo assembly. I have two MatePair files, one from Illumina MiSeQ and the other from IonTorrent. My problem is first how do i prepare the data before the assembly, and then, which assembler is better? (I have been working with Ray, MIRA, Newbler and SPAdes, these last two gave much better results in other data) In Illumina matepair, i tried to work with NxTrim but i need two files as an input and i do not know if a complementary file of the original will work, then i tried Trimmomatic too, and used these files as entrey (the original and the complementary i created) but i'm not sure if the output files will work for the assembler... Anyway, has anyone has any experience working with these kind of data? Which pipeline do you recommend? Thanks in advance.
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

    #2
    For reference cross-posted: https://www.biostars.org/p/191400/

    It is ok to cross-post on SeqAnswers.

    Comment

    • Markiyan
      Senior Member
      • Sep 2010
      • 126

      #3
      Try sff2phd (from consed contrib section) on ion's sff / miseq_trimmer on fastq....

      First you need to find out what are the naming rules for specifying (mate) pairs for your assembler of choice.
      Usually most assemblers are happy with [template_name]/1 for forward read and [template_name]/2 for reverse read name in fastq format. Some may want interleaved files and some may want forward/reverse reads in the separate files.

      Than we need to prepare the data.
      In any case the matepairs:

      >read(template)_name
      [seq/2C][linker][seq/1]

      have to be split:
      >read/1
      seq/1
      >read/2
      seq/2

      Than for SFF files (including iontorrent):
      get my sff2phd tool from the contrib section of the consed, update the linker sequence to the iontorrent's paired end one and run it (you can disable phd files generation, unless you would like to use phredPhrap assembly pipeline) and save fastq files.

      sff2phd produces fastq files with offset Q+64, while miseq_trimmer uses a standard fastq offset Q+31.

      For illumina files:
      1. run the input through the FLASH
      2. give an attached matepair splitter a try (at least on the overlapping ends file).

      fastq_miseq_trimmer [ext_frags.fastq] -base=[(filename_base).mpe1] -out1=[mpe1_1.fastq] -outlq=[mpe1_se.fastq] -out2=[mpe1_2.fastq] -mpestd -minl=32 -noclip -ends=3

      PS: If any improvements/fixes to the tools are required, feel free to contact me.
      PPS: Please can you post the linker sequence for the iontorrent PE reads, so I can add it to my tool.
      Attached Files
      Last edited by Markiyan; 05-17-2016, 04:51 AM. Reason: Attachments missing - fixxed.

      Comment

      Latest Articles

      Collapse

      • SEQadmin2
        Advanced Sequencing Platforms Tackle Neuroscience’s Toughest Genomics Problems
        by SEQadmin2



        Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
        ...
        07-09-2026, 11:10 AM
      • SEQadmin2
        Cancer Drug Resistance: The Lingering Barrier to Rising Survival
        by SEQadmin2



        Cancer survival rates have significantly increased in the last few decades in the United States, reaching a combined 70% 5-year survival rate by 2021. Behind this number, there are years of research to find new therapies, drug targets, and early detection methods. But there is one core challenge that keeps slowing down these advances, and it’s about drug resistance.

        There is no single reason why many patients don’t respond to treatment as expected. Cancer is...
        07-08-2026, 05:17 AM
      • 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

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by SEQadmin2, 07-13-2026, 10:26 AM
      0 responses
      23 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 07-09-2026, 10:04 AM
      0 responses
      33 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 07-08-2026, 10:08 AM
      0 responses
      20 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 07-07-2026, 11:05 AM
      0 responses
      34 views
      0 reactions
      Last Post SEQadmin2  
      Working...