Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Optimum insert size for De-novo transcriptome assembly

    I am making RNA-Seq libraries from mouse tissue for de-novo transcriptome assembly. I am planning to use cufflinks for this (the genome guided denovo assembly approach). What insert size is optimal for this? Since we are aligning to a well annotated genome, does it make sense to have a higher average insert size? This way, many more reads will span more than one exon with paired end sequencing, Won't this boost chances of finding alternative spliced isoforms? But does this entail sacrificing info on 3' and 5' end which is equally important (an important goal of our study is to find alternate 3' ends of genes)?
    Thanks for your inputs!

  • #2
    I think a broad range of insert sizes is good, particularly if you are interested in short transcripts that may not be represented at all if you filter out short inserts. The less size-selection, the less bias (aside from remove fragments that are too short to be useful or too long to amplify), and the greater the library uniqueness, which is important for assembly.

    Comment

    Latest Articles

    Collapse

    • seqadmin
      Essential Discoveries and Tools in Epitranscriptomics
      by seqadmin




      The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...
      04-22-2024, 07:01 AM
    • seqadmin
      Current Approaches to Protein Sequencing
      by seqadmin


      Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
      04-04-2024, 04:25 PM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by seqadmin, 04-11-2024, 12:08 PM
    0 responses
    59 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 10:19 PM
    0 responses
    57 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-10-2024, 09:21 AM
    0 responses
    51 views
    0 likes
    Last Post seqadmin  
    Started by seqadmin, 04-04-2024, 09:00 AM
    0 responses
    56 views
    0 likes
    Last Post seqadmin  
    Working...
    X