Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Produce different Tophat results with and without -z0 option

    Aligned paired-end RNA-seq using Tophat ver. 1.4.0.
    Used -z0 option for Tophat but it produced a different bam result.

    input fastq file size : 4.1G * 2 for paired-end
    ==================
    output 1. without -z0 option
    Code:
    $ tophat -r 94 -p 8 -G genes.gtf genome s_1_1.fastq s_1_2.fastq
    1.3G : accepted_hits.bam
    88K : deletions.bed
    50K : insertions.bed
    3.9M : junctions.bed
    70 : left_kept_reads.info
    4.0K : logs
    70 : right_kept_reads.info
    93M : unmapped_left.fq.z
    113M : unmapped_right.fq.z
    ==================
    output 2. with -z0 option
    Code:
    $ tophat -z0 -r 94 -p 8 -G genes.gtf genome s_1_1.fastq s_1_2.fastq
    523M : accepted_hits.bam
    55K : deletions.bed
    31K : insertions.bed
    2.6M : junctions.bed
    70 : left_kept_reads.info
    4.0K : logs
    70 : right_kept_reads.info
    2.7G : unmapped_left.fq
    2.5G : unmapped_right.fq
    ==================

    Then, I used samtools flagstat to look at bam file stats.
    The first bam file(without -z0 option) reported:
    Code:
    $ samtools flagstat accepted_hits.bam
    
    57149297 + 0 in total (QC-passed reads + QC-failed reads)
    0 + 0 duplicates
    57149297 + 0 mapped (100.00%:-nan%)
    57149297 + 0 paired in sequencing
    28845685 + 0 read1
    28303612 + 0 read2
    42295726 + 0 properly paired (74.01%:-nan%)
    54359924 + 0 with itself and mate mapped
    2789373 + 0 singletons (4.88%:-nan%)
    0 + 0 with mate mapped to a different chr
    0 + 0 with mate mapped to a different chr (mapQ>=5)
    The second bam file(with -z0 option) reported:
    Code:
    $ samtools flagstat accepted_hits.bam 
    
    20898015 + 0 in total (QC-passed reads + QC-failed reads)
    0 + 0 duplicates
    20898015 + 0 mapped (100.00%:-nan%)
    20898015 + 0 paired in sequencing
    10912428 + 0 read1
    9985587 + 0 read2
    3175304 + 0 properly paired (15.19%:-nan%)
    13747868 + 0 with itself and mate mapped
    7150147 + 0 singletons (34.21%:-nan%)
    0 + 0 with mate mapped to a different chr
    0 + 0 with mate mapped to a different chr (mapQ>=5)
    Does Tophat use different algorithm depend on -z0 option?
    How come Tophat produces different bam files?

    Thanks in advance!
    #####################
    # QUE LA FORCE SOIT AVEC TOI #
    #####################

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, 04-25-2024, 11:49 AM
0 responses
19 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-24-2024, 08:47 AM
0 responses
17 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-11-2024, 12:08 PM
0 responses
62 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 10:19 PM
0 responses
60 views
0 likes
Last Post seqadmin  
Working...
X