Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • Miseq assemblies

    Bacteria sequencing. Nextera XT library prep. Triplicate technical replicates from a single MiSeq run 2 x 250 PE, de novo assembly on MiSeq gives ~450 contigs for all three. Optimized velvet assembly from fastq's gives 200 contigs for two samples and 800 contigs for one. Why would there be so much of a difference between 3 technical replicates for one method of assembly and not for the other?
    To make things more complicated the triplicate samples are from a single strain but three individual DNA extractions.
    Last edited by RGLADSTONE; 01-11-2013, 02:28 AM.

  • #2
    Originally posted by RGLADSTONE View Post
    Bacteria sequencing. Nextera XT library prep. Triplicate technical replicates from a single MiSeq run 2 x 250 PE, de novo assembly on MiSeq gives ~450 contigs for all three. Optimized velvet assembly from fastq's gives 200 contigs for two samples and 800 contigs for one. Why would there be so much of a difference between 3 technical replicates for one method of assembly and not for the other?
    To make things more complicated the triplicate samples are from a single strain but three individual DNA extractions.
    1) is there a reference? If so, map and then estimate the insert size
    2) Try FLASH to see if the reads overlap
    3) Trim to 100bp PE and assemble again -> does this help?
    4) Randomly choose 10%, 25%, 50% of the data, assemble again, does this help?
    5) have you checked for adapters?

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Advancing Precision Medicine for Rare Diseases in Children
      by seqadmin




      Many organizations study rare diseases, but few have a mission as impactful as Rady Children’s Institute for Genomic Medicine (RCIGM). “We are all about changing outcomes for children,” explained Dr. Stephen Kingsmore, President and CEO of the group. The institute’s initial goal was to provide rapid diagnoses for critically ill children and shorten their diagnostic odyssey, a term used to describe the long and arduous process it takes patients to obtain an accurate...
      12-16-2024, 07:57 AM
    • seqadmin
      Recent Advances in Sequencing Technologies
      by seqadmin



      Innovations in next-generation sequencing technologies and techniques are driving more precise and comprehensive exploration of complex biological systems. Current advancements include improved accessibility for long-read sequencing and significant progress in single-cell and 3D genomics. This article explores some of the most impactful developments in the field over the past year.

      Long-Read Sequencing
      Long-read sequencing has seen remarkable advancements,...
      12-02-2024, 01:49 PM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, 12-17-2024, 10:28 AM
    0 responses
    25 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 12-13-2024, 08:24 AM
    0 responses
    42 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 12-12-2024, 07:41 AM
    0 responses
    28 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 12-11-2024, 07:45 AM
    0 responses
    42 views
    0 likes
    Last Post seqadmin  
    Working...
    X