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  1. IGB
  2. IGBF-3881

Add samtools filter by and color by options for CR and CB tags

    Details

    • Type: Task
    • Status: Closed (View Workflow)
    • Priority: Major
    • Resolution: Done
    • Affects Version/s: None
    • Fix Version/s: None
    • Labels:
      None

      Description

      For this ticket, explore the prospect of adding samtools tags to right-click filter function so that user can filter reads from the track based on samtools tags. For the purposes of this ticket, adding CR, cellular barcode sequence bases, as a filter option so that users can filter reads based on cell identifiers.

      Adding this filter may help in understanding the UMI count matrix and to make a plan to add features for single cell data.

        Attachments

        1. After Color by CR.png
          73 kB
          Karthik Raveendran
        2. After Filter by CR.png
          16 kB
          Karthik Raveendran
        3. Before Color by CR.png
          23 kB
          Karthik Raveendran
        4. Before Filter by CR.png
          20 kB
          Karthik Raveendran

          Issue Links

            Activity

            Hide
            karthik Karthik Raveendran added a comment -

            Completed implementing filter by samtools tags options. We can filter reads by CR number which is cell barcode number. Working on the color by option.

            Show
            karthik Karthik Raveendran added a comment - Completed implementing filter by samtools tags options. We can filter reads by CR number which is cell barcode number. Working on the color by option.
            Show
            karthik Karthik Raveendran added a comment - Color by commit: https://bitbucket.org/KarthikRavee91/karthikfork-igb/commits/771e8ab7b8a4f268bbe249a65a96aa3146703c9f Filter by commit: https://bitbucket.org/KarthikRavee91/karthikfork-igb/commits/771e8ab7b8a4f268bbe249a65a96aa3146703c9f
            Hide
            karthik Karthik Raveendran added a comment -

            During a demo showing Nowlan Freese the new filter and color options there was an error which I was not able to replicate after that. Probably someone else testing it would be better. Including the link to download installer page in my repo: https://bitbucket.org/KarthikRavee91/karthikfork-igb/downloads/

            Show
            karthik Karthik Raveendran added a comment - During a demo showing Nowlan Freese the new filter and color options there was an error which I was not able to replicate after that. Probably someone else testing it would be better. Including the link to download installer page in my repo: https://bitbucket.org/KarthikRavee91/karthikfork-igb/downloads/
            Hide
            karthik Karthik Raveendran added a comment - - edited

            Testing Protocol

            Prepare for Test:

            1. Go to https://bitbucket.org/KarthikRavee91/karthikfork-igb/downloads/ and download the IGBF-3881 installer relevant to your OS
            2. Install and open that IGB
            3. Download the quickload folder https://drive.google.com/drive/folders/14LRo1FQ0daM0oZKZ17bo8tuW3qv0GAKk?usp=drive_link
            • Add the quickload to Data Sources as " 1k A375 "
            • Press Alt+P and click on the Data Sources tab
            • Click on Add
            • Add the path of the quickload you just downloaded
            • Use " 1k A375 " as the server name
            1. Click on Homo Sapiens in the carousel to load the human reference genome and click to select the track
            2. Go to the annotation tab in the bottom panel of IGB
            3. In the track height section, check Lock Track height and set height to 150
            4. Go Data Access tab and expand 1k A375 folder on Available Data section
            5. Check the only file under that folder
            6. Go to the search bar on top left and search for DPH2
            7. Click on Load Data and Load Sequence (top-right)
            8. Right-click on one of the reads and click on Get Info
            9. In the window that appears, scroll to the key "CR" and copy the value and close window

            Feature Test #1

            1. Right-click on the 1k A375 track and click on "Color By"
            2. Click OK
            3. Select Samtools Tags in Color By dropdown
            4. Paste the CR value in the value text box
            5. Click OK

            Expected Observation: Reads with the CR value that you copied turns green and maybe some other reads as well and everything else turns red. Please check if the green ones have the same CR value as the one that you copied.

            Feature Test #2

            1. Right-click on the 1k A375 track and click on "Filter..."
            2. Click on add button
            3. Select Samtools Tags in Show only
            4. Paste the CR value in the value text box and select "=" in the comparator dropdown
            5. Click OK

            Expected Observation: Except for the reads with the CR value that you copied, everything else disappears. Please check if the ones that remain have the same CR value as the one that you copied.

            Show
            karthik Karthik Raveendran added a comment - - edited Testing Protocol Prepare for Test: Go to https://bitbucket.org/KarthikRavee91/karthikfork-igb/downloads/ and download the IGBF-3881 installer relevant to your OS Install and open that IGB Download the quickload folder https://drive.google.com/drive/folders/14LRo1FQ0daM0oZKZ17bo8tuW3qv0GAKk?usp=drive_link Add the quickload to Data Sources as " 1k A375 " Press Alt+P and click on the Data Sources tab Click on Add Add the path of the quickload you just downloaded Use " 1k A375 " as the server name Click on Homo Sapiens in the carousel to load the human reference genome and click to select the track Go to the annotation tab in the bottom panel of IGB In the track height section, check Lock Track height and set height to 150 Go Data Access tab and expand 1k A375 folder on Available Data section Check the only file under that folder Go to the search bar on top left and search for DPH2 Click on Load Data and Load Sequence (top-right) Right-click on one of the reads and click on Get Info In the window that appears, scroll to the key "CR" and copy the value and close window Feature Test #1 Right-click on the 1k A375 track and click on "Color By" Click OK Select Samtools Tags in Color By dropdown Paste the CR value in the value text box Click OK Expected Observation: Reads with the CR value that you copied turns green and maybe some other reads as well and everything else turns red. Please check if the green ones have the same CR value as the one that you copied. Feature Test #2 Right-click on the 1k A375 track and click on "Filter..." Click on add button Select Samtools Tags in Show only Paste the CR value in the value text box and select "=" in the comparator dropdown Click OK Expected Observation: Except for the reads with the CR value that you copied, everything else disappears. Please check if the ones that remain have the same CR value as the one that you copied.
            Hide
            pkulzer Paige Kulzer added a comment - - edited

            Thank you for the detailed protocol, Karthik Raveendran! This worked really well!

            I walked through all of the steps outlined above and got the expected results as described for each Feature Test. I also zoomed out a bit from the DPH2 gene and made sure that the behavior I was seeing at that locus was occurring elsewhere in previously unloaded parts of the genome, too.

            For reference, the CR value I was using was: ACGAAAGGTGCTTCCG
            And the name of the read I initially clicked on to get that value was: A00984:1229:HTJHLDRX3:2:2137:16938:4773

            Show
            pkulzer Paige Kulzer added a comment - - edited Thank you for the detailed protocol, Karthik Raveendran ! This worked really well! I walked through all of the steps outlined above and got the expected results as described for each Feature Test. I also zoomed out a bit from the DPH2 gene and made sure that the behavior I was seeing at that locus was occurring elsewhere in previously unloaded parts of the genome, too. For reference, the CR value I was using was: ACGAAAGGTGCTTCCG And the name of the read I initially clicked on to get that value was: A00984:1229:HTJHLDRX3:2:2137:16938:4773
            Hide
            karthik Karthik Raveendran added a comment -

            Submitting a pull request on this branch is put on hold until the release branch is created for the Release IGB 10.1.0.

            Show
            karthik Karthik Raveendran added a comment - Submitting a pull request on this branch is put on hold until the release branch is created for the Release IGB 10.1.0.
            Show
            karthik Karthik Raveendran added a comment - PR Submitted: https://bitbucket.org/lorainelab/integrated-genome-browser/pull-requests/1047
            Hide
            nfreese Nowlan Freese added a comment -

            Tested with main branch installer on Mac using Karthik's instructions above. Everything working correctly, no errors in the logs.

            Closing ticket.

            Show
            nfreese Nowlan Freese added a comment - Tested with main branch installer on Mac using Karthik's instructions above. Everything working correctly, no errors in the logs. Closing ticket.

              People

              • Assignee:
                karthik Karthik Raveendran
                Reporter:
                karthik Karthik Raveendran
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                Dates

                • Created:
                  Updated:
                  Resolved: