Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Cuffmerge Error

    I am in the process of analysis my first RNA-seq experiment and have ran into an error with Cuffmerge that I just can't solve. I've read the threads of people who have had the same error and it hasn't seemed to help. So far I've aligned my reads using STAR-2.5.2b and used these to run Cufflinks with the command,

    Code:
    ~/jgg-093-aa/cufflinks-2.2.1.Linux_x86_64/cufflinks \
    	--library-type fr-firststrand -p 8 -G ~/jgg-093-aa/star/genome/genes.gtf \
    	-o ~/jgg-093-aa/Cufflinks/Treatment1/ \
    	~/jgg-093-aa/Aligned_Rn6/Treatment1Aligned.sortedByCoord.out.bam
    This created a folder with the transcripts.gtf file for all of my samples. I am now trying to merge all these GTF files with Cuffmerge before going on to use Cuffdiff.

    My Cuffmerge script that I have been trying to submit is:

    Code:
    ~/jgg-093-aa/cufflinks-2.2.1.Linux_x86_64/cuffmerge \
    	-g ~/jgg-093-aa/star/genome/genes.gtf -o ~/jgg-093-aa/Cufflinks/Master/  -s ~/jgg-093-aa/star/genome/Rattus_norvegicus/UCSC/rn6/Sequence/WholeGenomeFasta/genome.fa \
    	~/jgg-093-aa/Cufflinks/Assemblies.txt
    Every time I try running this though, I get the following output:

    Code:
    [Thu Mar 23 17:05:51 2017] Beginning transcriptome assembly merge
    -------------------------------------------
    
    [Thu Mar 23 17:05:51 2017] Preparing output location /home/rmarti68/jgg-093-aa/Cufflinks/Master//
    Traceback (most recent call last):
      File "/home/rmarti68/jgg-093-aa/cufflinks-2.2.1.Linux_x86_64/cuffmerge", line 580, in <module>
        sys.exit(main())
      File "/home/rmarti68/jgg-093-aa/cufflinks-2.2.1.Linux_x86_64/cuffmerge", line 538, in main
        gtf_input_files = test_input_files(transfrag_list_file)
      File "/home/rmarti68/jgg-093-aa/cufflinks-2.2.1.Linux_x86_64/cuffmerge", line 268, in test_input_files
        g = open(line,"r")
    IOError: [Errno 2] No such file or directory: '~/jgg-093-aa/Cufflinks/Treatment1/transcripts.gtf'
    I've doubled checked that the path to the transcripts.gtf file is correct and using the paths for the other components with Cufflinks and STAR worked.

    Any help or advice is very appreciated.

    Thanks!
    Attached Files

Latest Articles

Collapse

  • 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
  • seqadmin
    Strategies for Sequencing Challenging Samples
    by seqadmin


    Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
    03-22-2024, 06:39 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by seqadmin, 04-11-2024, 12:08 PM
0 responses
23 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 10:19 PM
0 responses
24 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-10-2024, 09:21 AM
0 responses
20 views
0 likes
Last Post seqadmin  
Started by seqadmin, 04-04-2024, 09:00 AM
0 responses
52 views
0 likes
Last Post seqadmin  
Working...
X