Seqanswers Leaderboard Ad

Collapse

Announcement

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

  • Generating SFF files

    By mistake the SFF file was deleted once the Amplicon Full processing pipeline was done. How can we reprocess the data to generate the SFF file back again?
    Thanks!

  • #2
    Originally posted by Xterra View Post
    By mistake the SFF file was deleted once the Amplicon Full processing pipeline was done. How can we reprocess the data to generate the SFF file back again?
    Thanks!
    Sadly the gsReporter application will not produce SFF files from an exiting analysis directory; SFF files can only be generated by gsRunProcessor. But you don't have to repeat the entire Signal Processing pipeline, you can perform an Analysis Filter job on the existing D_ directory which will take far less time.

    If you are unfamiliar with how to perform a Filter only analysis (these steps assume your current working directory to be the run (R_) directory and the Full Amplicon analysis directory (D_) still exists there):

    1. Create a template file with the default amplicon filter settings, saving it to a file named (e.g.) ampliconFilter.xml:

    Code:
    # gsRunProcessor --template=filterOnlyAmplicon > ampliconFilter.xml
    2. Run the filter only analysis pipeline providing the name of the template file you just generated and the existing analysis (D_) direcotry.

    Code:
    # runAnalysisFilter --pipe=./ampliconFilter.xml <D_directory_name>
    The filter only pipeline runs much faster than the full analysis pipeline. You will have a new, complete D_ directory with the SFF files.

    Comment


    • #3
      kmcarr

      I actually did exactly what you suggested but the new D directory did not included the SFF file on the regions subfolder.

      Comment


      • #4
        Originally posted by Xterra View Post
        I actually did exactly what you suggested but the new D directory did not included the SFF file on the regions subfolder.
        The SFF files are not in the regions subfolder. There should be a separate subdirectory named 'sff'.

        Comment


        • #5
          kmcarr

          Sorry that's what I meant. Now, I am not quite sure what's going on, I repeated the process three time and I finally got the SFF file. However, when I split the MID, I can see I got very few reads/MID. When I split the original SFF file that came out of the run, I had plenty of reads per MID. Therefore, something is wrong.
          Now, I can see the fasta and qual files, can I split the MID from the fasta file using any of the SFF tools?
          Last edited by Xterra; 10-31-2011, 08:19 AM.

          Comment


          • #6
            Originally posted by Xterra View Post
            Sorry that's what I meant. Now, I am not quite sure what's going on, I repeated the process three time and I finally got the SFF file. However, when I split the MID, I can see I got very few reads/MID. When I split the original SFF file that came out of the run, I had plenty of reads per MID. Therefore, something is wrong.
            Now, I can see the fasta and qual files, can I split the MID from the fasta file using any of the SFF tools?
            Why do want to do that? You should investigate on what is going wrong with your filtering job ...

            just my 2p,
            Sven

            Comment


            • #7
              At this point I am more interested on retrieving my data.
              Do you know of any tool I can use to split the MID using the central FASTA file that was generated during the run?

              Comment


              • #8
                You finally will need to find out what was wrong, believe me ... :-)

                Nevertheless, one of the tools capable of doing what you want might be "FASTX Barcode Splitter" from the fastx toolkit (http://hannonlab.cshl.edu/fastx_tool...splitter_usage).

                hth, Sven

                Comment


                • #9
                  sklages

                  Thanks! I will check it out.

                  Comment

                  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
                  22 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
                  19 views
                  0 likes
                  Last Post seqadmin  
                  Started by seqadmin, 04-04-2024, 09:00 AM
                  0 responses
                  50 views
                  0 likes
                  Last Post seqadmin  
                  Working...
                  X