Mission Bio's Tapestri Pipeline allows customers to process single-cell DNA and DNA + Protein sequencing data generated on the Tapesti Platform.

Create a Tapestri Pipeline Account and Launch Tapestri Pipeline

Create an Account

If you are not an active Mission Bio customer, you must first create a Mission Bio Portal account. To create a Mission Bio account, enter your email address and a password with these requirements on Tapestri Portal. If you have problems creating an account, see this article.

Tapestri Pipeline is only available to customers. If the Launch button next to Tapestri Pipeline on Tapestri Portal is gray, click How do I get access?, and a dialog box displays that submits your request for access. Mission Bio will create a new Tapestri Pipeline account free of charge if you purchased the Tapestri Platform. Please contact sales@missionbio.com for purchasing information.

Once created, you will receive an email to set up your account.

If you do not receive a confirmation email, check your spam folder. If it is not there, contact support@missionbio.com.

Managing Users in Your Group

If you are the Admin for your Tapestri Pipeline account, you can add and remove members from your group. Go to the menu in the upper-right corner in Tapestri Portal and select Groups.

Add Members

1. Click the edit users icon under the Actions column.
2. Click Add Member.
   
   
3. Add the email ids for the users you want to add, separate them with either a comma (,) or semicolon (;). Select the role.
   Notes
   1. You can only add 1 user type at a time.
   2. You must enter either a "," or ";" after each email, even if there is only one address. The email address will turn blue.
      
      
      
4. Click Add Member(s).
   An email is sent to each email address added. Once an invitation is accepted, the member will be listed in the Members tab. Until then, they will be in the Invited tab.
   

Remove Members

1. Click the trash can icon in the Actions column. It will automatically delete the member without a warning message. You can delete Members and pending invitations.

Supported Browsers

Chrome v37+

Edge v80+

Firefox v41+

Opera

Safari v10.2+

Launch Tapestri Pipeline

Launch Tapestri Pipeline from Tapestri Portal.

Settings Menu

In the upper-left corner, click the settings menu.

Home

Clicking Home takes you to the Runs page.

My Account

Clicking My Account takes you to the Profile Details page where you can update your contact information and change your password.

Cloud Connector

Tapestri Pipeline v2 allows importing files from Illumina BaseSpace and Amazon S3. To use either of these options, first set up the Cloud Connectors.

Illumina BaseSpace

To generate a BaseSpace access token, follow these instructions.

Amazon S3

To learn about how Tapestri Pipeline integrates with Amazon S3 buckets, read this FAQ.

Support

To view the User Guide, click the Support menu item.

Logout

To log out of Tapestri Pipeline, click the Logout menu item.

Dashboard

The dashboard is the navigation section at the top of the Runs and Files pages.

In-Progress, Failed, Completed

Depending on whether the Runs or Files page is displayed, the dashboard displays the total number of runs/files In-Progress, Failed, and Completed. The information contained in this section pertains to the previous 30 days.

Refresh, Add Files, Start Run

Refresh

Use the refresh button to update the status in the Runs or Files table, depending on the displayed page.

Add Files

To add files, click this button. For more information, see this section.

Start Run

To start a run, click this button. For more information, see this section.

Runs Page

Selecting the Runs page from the top-left corner displays a table with all the runs. It displays the following information:

• Run Name
• Pipeline
• Status
• Duration
• Start Date
• User

This additional information is displayed for DNA and DNA + Protein runs.

• Cell Count
• Panel Uniformity
• Mean Reads/Cell/Amplicon
• % DNA Read Pairs Assigned to Cells
• Mean Reads/Cell/Antibody (DNA + Protein only)

Search

To search for specific runs, type the criteria into the search box. You can search for:

• Run Names
• DNA + Protein
• Status
• User

Sort

Sort the table by clicking any column name. It sorts in both ascending and descending order. Click it again to sort the other way.

Cancel Run

Click the icon to the left of the Run Name to Cancel Run. This only applies to in-progress runs.

Clone Run

Click the icon to the left of the Run Name to Clone Run. Only completed or canceled DNA and DNA + Protein runs can be cloned. It will prompt you to Name the Run. After clicking Proceed, the parameters of the run can be changed.

Delete / Rename Run

Runs cannot be deleted or renamed once submitted.

Run Details

Clicking the Run Name displays the Run Details page for that run. See this section for details.

Files Page

Clicking the Files option from the top-left corner displays a table with all the files in sections for FASTQ and Panel files.

The table displays the following information for each file type:

• File Name
• File Type
• Status
• Uploaded Date
• File Size
• Uploaded By
• Source

Search

To search for specific files, type the criteria into the search box. You can search for:

• File Name
• File Type
• Status
• Uploaded By
• Source

Sort

Sort the table by clicking any column name. It sorts in both ascending and descending order. Click it again to sort the other way.

Delete / Rename / Cancel File

Click the icon to the left of the File Name of a completed file to Delete or Rename the file.

Click the icon to the left of the File Name of an in-progress file to Cancel the upload. It will also cancel all other files that were uploaded at the same time.

Note: You cannot delete or rename the Catalog panels uploaded by Mission Bio – AML, Myeloid, CLL, THP, and TotalSeq™-D Heme Oncology Cocktail.

File Type

Click the File Type popup to change the file type.

Starting a Run

There are two ways to start a run – Start Run and Add Files then Start Run.

1. Start Run

Click the Start Run button in the top-right corner.

Name the Run

• Add a run name. Since this is used as the prefix for the output files, it cannot contain spaces or special characters, such as % & * @ ^ ; “, ' *.
• Add an optional description.
• Click Next.

Step 01 – Select Pipeline & Other Parameters

• Select the Pipeline
  Choose DNA, DNA + Protein, or Merge Samples in the dropdown menu.
  DNA: This requires a DNA Panel file and an even number of DNA FASTQ files.
  DNA + Protein: This requires a DNA Panel file and an even number of DNA FASTQ files and a Protein Panel file and an even number of Protein FASTQ files. The number of DNA and Protein FASTQ files must be the same.
  Merge Samples: This requires at least two runs to merge. While the number of runs to merge is currently unlimited, Mission Bio recommends only merging 2 to 5 runs. One .h5 file will be created. When merging DNA and DNA + Protein runs, only the DNA information is retained. Note: Ensure that the merged run name is unique.
  
  
• Select the Reference Genome (DNA, DNA + Protein)
  Choose either Human or Mouse in the dropdown menu.
  
  
• Generate cells.bam file only (DNA only)
  This stops the run after generating the cells.bam file. This sequence alignment file contains only the reads that belong to cells and can be used for genotyping with any tool.
  
  
• Add Panel Files (DNA, DNA + Protein)
  Choose either Select from Existing Files or Upload from Local Computer in the File Source dropdown menu. Click the Add button.
  
  File Format
  DNA Custom Panel File: A .zip file, Upload from Local Computer
  DNA Designer Catalog Panel File: 4 Preloaded, Select From Existing Files
  Protein Custom Panel File: A .csv file, Upload from Local Computer
  Protein Designer Catalog Panel File: 1 Preloaded, Select From Existing Files
  
  Number of Files
  DNA: Select or upload one DNA panel file.
  DNA + Protein: Select or upload one DNA and one Protein Panel file.
  
  
• Select Runs To Merge (Merge Samples only)
  Select which runs to merge.
  
  
• Click Next.

Step 02 – Select the FASTQ Files (DNA, DNA + Protein)

For DNA runs, select an even number of DNA FASTQ files.
For DNA + Protein runs, select an even number of DNA and Protein FASTQ files. The number of DNA and Protein FASTQ files must be equal.

File Format
Only files with these extensions are allowed: .fastq; .fastq.gz; .fq; and .fq.gz. When importing files from Amazon S3 folders, all the files in the folder and nested folders with the correct extensions will import. All others will be ignored.

To use Amazon S3 or Illumina BaseSpace, first set up the Cloud Connector.

• In the dropdown, choose Select from Existing Files, Upload from Local Computer, Import from Amazon S3, or Import from Illumina BaseSpace in the Select FASTQ Files dropdown menu.
  
  
• Click the Add button.
  
  1. Select from Existing Files
  Click the checkbox to the left of all the files you want to include.
  Click Done.
  
  2. Upload from Local Computer
  DNA
  Drag and drop the DNA FASTQ files into the upload area or click Choose Files.
  
  DNA + Protein
  Drag and drop the DNA FASTQ files into the upload area or click Choose Files.
  Choose Protein FASTQ from the dropdown menu, and drag and drop the Protein FASTQ files into the upload area or click Choose Files.
  
  Click Upload.
  
  3. Amazon S3
  To add multiple files or folders, separate the names with either a "," or ";".
  
  DNA
  Add the Amazon S3 URI for the DNA FASTQ files or the folder containing the FASTQ files.
  
  DNA + Protein 
  Add the Amazon S3 URI for the DNA FASTQ files or the folder containing the FASTQ files.
  Click +Add FASTQ files for another analyte.
  Select Protein FASTQ.
  Add the Amazon S3 URI for the Protein FASTQ files or the folder containing the FASTQ files.
  
  Click Import.
  
  4. Illumina BaseSpace
  To add multiple biosamples, separate the names with either a "," or ";".
  
  DNA
  Add the Biosample Name for the DNA FASTQ files with this format – Project ID: Biosample name.
  
  DNA + Protein 
  Add the Biosample Name for the DNA FASTQ files – Project ID: Biosample name.
  Click +Add FASTQ files for another analyte.
  Select Protein FASTQ.
  Add the Biosample Name for the Protein FASTQ files – Project ID: Biosample name.
  
  Click Import.
  
  
• Click the Next button.

Step 03 – Lane Assignment (DNA, DNA + Protein)

Tapestri Pipeline auto assigns FASTQ files to the lanes. If you prefer to assign the files yourself, click the Reset button in the left panel. The files will then go into the right panel where you can drag and drop them into the correct lane location. Each lane should have 2 files, one R1 and one R2. You can have up to 4 lanes per run.

Note: Depending on the chemistry used to generate the FASTQ files, the sequencing library may have 1 or more pairs of FASTQ files in a run. If this is the case, there will a dropdown with the number of pairs instead of the static 1 in the image above. Please review the lane assignment rules in the User Guide.

Red and Green Highlighting

The green color indicates that the files follow the lane assignment rules we expect. For example, L001 indicates Lane 1. R1 denotes R1.

If the color is red, the file does not match our lane assignment rules. You can submit the run, but double-check to see that the files are in the correct location. Please review the lane assignment rules in the User Guide.

• Verify the file assignments for the DNA FASTQ files and Protein FASTQ files, when applicable.
• Click the Next button.

Step 04 – Preview

This is the final step before submitting the run. Confirm all the information. To edit any of the information, click the Edit icon next to the Steps or click the Step in the left Panel.

After confirming all the information, click Submit Run.

Run Successfully Submitted

Once the run is submitted, you have the option to Submit Another Run or Go To Runs Page.

Submit Another Run

This option clones the current run. Name the run and adjust any of the parameters.

Go To Runs Page

This option returns you to the main Runs page.

2. Add Files then Start Run

This option allows you to upload files before starting a run.

• Click Add Files.
  There are three ways to upload FASTQ files – Upload from Local Computer, Import from Amazon S3, and Import from Illumina BaseSpace. Panel files can only be uploaded from a local computer.
  
  
  
• For instructions on uploading files, refer to this section.

Click the Start Run button.

Follow these instructions to start the run.

Lane Assignment Rules

Tapestri Pipeline auto assigns files to lanes based on our naming convention. If the file names agree with our naming convention, then they are highlighted in green when assigned to lanes. If they do not conform, then they have a red background. This does not mean they are incorrect, however. For example, some people merge runs, which creates a Lane 5. Since Tapestri Pipeline only allows 4 lanes, this does not follow our conventions, thus a red highlight.

To determine lane assignment, we use the file name, so the file name should contain _R1_ and _R2_ for the algorithm to work. We parse the file name to determine the prefix. For the file names DNA-test_L001_R1_001.fastq.gz and DNA-test_L001_R2_001.fastq.gz, we consider “DNA-test” the prefix and then assign files to lane1 (L001).

Other rules:

1. 1 pair of FASTQ files
   Assigned to Lane1.
   Example: DNA-test_L001_R1_001.fastq.gz, DNA-test_L001_R2_001.fastq.gz
   
   
   
2. More than 1 pair of FASTQ files
   1. Same prefixes and extensions
      If you have a single library sequenced on multiple lanes of the flow cell, you may have 2 to 4 pairs of files. These files typically have the same prefixes and same extensions with different lane numbers, like L001 and L002. These files will be assigned to different lanes.
      
      
      
   2. Different prefixes
      There are 2 scenarios where you have files with different prefixes in a run:
      1. Technical replicates
         If you sequenced your sample twice to increase coverage, you will have 2 pairs of files with the same or different names. To analyze these samples, add them to 2 lanes on the first page. This scenario will be highlighted red, but ignore this.
         
      2. Tapestri Pipeline v1
         When working with an older Tapestri platform (V1 sequencing chemistry), your library will generate 2 or more pairs of FASTQ files – 2 when you use a single lane and 2+ for multiple lanes of a flow cell. In this case, the files are assigned to 2 sections as seen in the dropdown at the top of the left panel. 
         

Follows our convention for Lane Assignments

When the file names follow our convention, they have a green highlight. File names with Read Numbers (_R1_ and _R2_) and Lane Numbers (L001, L002, L003, and L004) abide by the rules. For example:

DNA-test_L001_R1_001.fastq.gz, DNA-test_L001_R2_001.fastq.gz

Protein-test_L001_R1_001.fastq.gz, Protein-test_L001_R2_001.fastq.gz

Does not follow the convention

These file names do not follow our convention, but you can proceed with some but not others. Each case below results in a red highlight. To rename a file, refer to this section.

1. Scenarios that will cause the pipeline to fail
   1. No R1 or R2 read in the run
      DNA-test_L001_R1_001.fastq.gz, DNA-test_L001_R1_001.fastq.gz
   2. Files from different lanes added to the same lane
      DNA-test_L001_R1_001.fastq.gz, DNA-test_L004_R2_001.fastq.gz 
2. Scenarios that will not cause the pipeline to fail, if you know the files are correct
   1. A mismatch in the lane number in the lane assignment and the lane number in the file name. In this case, ensure R1 and R2 have the same lanes.
      Protein-Test_L005_R1_001.fastq.gz, Protein-test_L005_R2_001.fastq.gz
   2. A mismatch in the prefixes. Confirm the files are correct before proceeding.
      Test_L001_R1_001.fastq.gz, Protein-test_L001_R2_001.fastq.gz
   3. Missing R1 and R2 in the filename. It is good practice to have these in the file name, but it will not cause the pipeline to fail. Confirm the lane assignment is correct.
      Protein-test_L005_001.fastq.gz, Protein-test_L005_001.fastq.gz
      

Run Report

For detailed information on the items contained in the Report, see this article. To download the Run Report, go to the Output Files tab and download the <fileprefix>.report.html file.

The DNA-only runs with "Generate cells.bam file only" and Merge Samples runs do not generate a Run Report.

Summary

The Summary page displays the following information:

• # cells
• Panel uniformity
• Mean reads/cell/amplicon
• % DNA read pairs assigned to cells
• Mean reads/cell/antibody
• Sample information
  • Run ID
  • Analyte
  • DNA panel name
  • DNA panel size
  • Reference genome
  • Chemistry version
  • DNA pipeline version
  • Protein panel name
  • Protein panel size
  • Protein pipeline version
  • Date analyzed
• Sequencing (Protein)
  • # total read pairs
  • Read quality (Q30)
  • % read pairs trimmed
  • % read pairs after cell barcode processing
  • % read pairs after antibody barcode processing
  • # total barcodes
  • # candidate barcodes
  • % read pairs after candidate barcode filtering
• Antibody counting
  • Mean reads/cell/antibody
  • # antibodies detected
  • Median antibodies/cell
• Panel uniformity plot
  • Normalized read counts (x-axis) per Amplicon (y-axis)
• Sequencing (DNA)
  • # total read pairs
  • Read quality (Q30)
  • % read pairs trimmed
  • % read pairs with valid barcodes
  • # total barcodes
• Mapping
  • % reads mapped to genome
  • % reads mapped to target
• Cell calling
  • # cells
  • Panel uniformity
  • Mean reads/cell
  • Mean reads/cell/amplicon
  • % DNA read pairs assigned to cells
  • DNA data completeness
  • % antibody read pairs assigned to cells
• Variant calling
  • # filtered variants
  • ADO rate

Advanced

The Advanced page displays the following information:

• Panel performance
  • % amplicons between 0.2*mean and 5*mean reads/cell/amplicon
  • % amplicons between 0.5*mean and 2*mean reads/cell/amplicon
  • % amplicons > 1x coverage
  • % amplicons > 5x coverage
  • % amplicons > 10x coverage
  • % amplicons > 20x coverage
  • % amplicons > 40x coverage
  • % reads to amplicons above 2*mean reads/cell/amplicon
• Low-performing amplicons
  • Amplicon name with mean read
• Amplicons with > 2* standard deviation Gini score
  • List of amplicons by name
• Amplicons with R1R2 imbalance
  • List of amplicons by name
• ADO calculation summary
• Amplicon Gini score distribution plot
  • Gini score (x-axis) by Number of amplicons (y-axis) with mean and threshold

Diagnostics

The Diagnostics page displays the following plots:

• R1R2 imbalance plot
  • x-axis: R1/R2 read percents
  • y-axis: Amplicons
  • Legend: R1 read percent, R2 read percent
• Balance R1R2 barplot
  • x-axis: R1/R2 read percents
  • y-axis: Amplicons
  • Legend: R1 read percent, R2 read percent
• DNA vs Protein reads scatter plot (Protein only)
  • x-axis: DNA reads
  • y-axis: Protein reads
  • Legend: Cell, Not cell
• Antibody cell distribution plot (Protein only)
  • x-axis: Antibodies
  • y-axis: Number of cells
• Read log-log plot
  • x-axis: Rank-ordered barcodes
  • y-axis: Total reads
  • Legend: DNA, Protein, Both
• DNA 1x vs 10x coverage plot
  • x-axis: Coverage at 1x
  • y-axis Coverage at 10x
  • Legend: Cell, Not cell, Unity line
• Antibody read distribution plot (Protein only)
  • x-axis: Antibodies
  • y-axis: log10(1 + number of antibody reads)

Output Files

The following output files are created based on the Pipeline.

DNA  and DNA + Protein

• all.barcode.cell.distribution.merged.tsv (DNA, DNA + Protein)
  This file reports the number of forward reads assigned to each amplicon for each cell found in the run. For more information, see this file.
  
  
• allele.drop.out.report.txt (DNA, DNA + Protein)
  This text file lists the variants used to calculate the allele dropout (ADO) rate. It shows the summary for the ADO calculation, such as the sample name, median ADO for the sample, number of variants, and total number of cells.
  
  Refer to this article to learn more about ADO calculation.
  
  
• barcode.cell.coverage.tsv (DNA, DNA + Protein)
  This file lists each amplicon and the number of mean reads per cell. For each amplicon, it also lists whether the amplicon passed the threshold. The threshold calculation is the mean of the total mean reads, e.g., the mean of column B * 0.2. If the mean reads for an amplicon is above this number, it passed the threshold and has a value of TRUE. If it is below this number, the value is FALSE.
  
  
• barcode.cell.distribution.merged.tsv (DNA, DNA + Protein)
  A file that reports the number of forward reads assigned to each amplicon for each cell found in a run. For more information, see this file.
  
  
• cells.bam (DNA, DNA + Protein)
  This file lists a read group (RG) tag for each read, which can be used to find the number of reads for the barcode. If your run is from Tapestri v1 using v1 chemistry, you will have multiple cells.bam files because v1 was index-specific.
  
  A .bam file is a binary version of a sequence alignment map (SAM). For more information on these formats, refer to this PDF. To visualize these files, use tools like IGV. 
  
  
• cells.bam.bai (DNA, DNA + Protein)
  This is the .bam index file used by the IGV tool to view the alignments.
  
  
• cells.loom
  This file is an input file for Tapestri Insights v2. LOOM is an efficient file format for very large omics datasets, consisting of a main matrix, optional layers, and a variable number of row and column annotations. For more information, see this file.
  
  
• cells.vcf.gz (DNA, DNA + Protein)
  This compressed annotated .vcf file conforms to the standard GATK format. It contains all the variants for all the barcodes called as cells. 
  
  
• dna.h5 (DNA only) / dna+protein.h5 (DNA + Protein only)
  
  The .h5 files are the input to Tapestri Insights v3. It is a new multi-omics file format that contains the data for all the analytes in a single file. For more details, read this article.
  
  
• fastp.html (DNA, DNA + Protein)
  This .html report file is generated by the FASTP tool that is used to check the input sequence quality. It reports various metrics and graphs, such as base quality – Q20 and - Q30 base count and base content of the reads.
  
  
• flagstat.txt
  This text file shows the mapping statistics from the reference genome alignment step, listing the number of QC-passed reads and QC-failed reads for items like read1, read2, properly paired, and mapped. If your run is from Tapestri v1 using v1 chemistry, you will have multiple flagstat.txt files because v1 was index-specific. For more information on the flagstat format, see this description.
  

• mapped.bam (DNA, DNA + Protein)
  This file is generated after mapping to the reference genome and selecting high-quality primary alignments with a mapping quality score of > 30. It is generated after barcode correction and alignment but before cell calling. It maps all the barcodes in the run. If your run is from Tapestri v1 using v1 chemistry, you will have multiple mapped.bam files because v1 was index-specific.
  
  A .bam file is a binary version of a sequence alignment map (SAM). For more information on these formats, refer to this PDF. To visualize these files, use tools like IGV.

• metrics.json (DNA only)
  This file contains the .HTML run report content in the .json format. It is a machine-readable format and provides an easy way to do additional analysis on the run report metrics.
  
  
• report.html (DNA, DNA + Protein)
  This new .html report is generated for each run. It summarizes the run details and all the information that was part of the other output files in Tapestri Pipeline v1. For more details, see this section.
• report.json (DNA + Protein only)
  This file contains the .HTML run report content in the .json format. It is a machine-readable format and provides an easy way to do additional analysis on the run report contents.

Merge Samples

The Merge Samples Pipeline generates a single merged .h5 file that contains different assays, depending on the analytes present in the input runs. 

1. Merge 2 or more DNA-only runs – Merged .h5 file with only the DNA-related assays – dna_variants and dna_read_counts.
2. Merge 2 or more DNA + Protein runs – Merged .h5 file with DNA and protein assays – dna_variants, dna_read_counts, and protein_read_counts.
3. Merge DNA and DNA + Protein runs – Merged .h5 file with only the DNA-related assays – dna_variants and dna_read_counts.

Sort

Sort the table by clicking any column name. It sorts in both ascending and descending order. Click it again to sort the other way.

 

Download

To download any output file, click the download icon to the left of the File Name.

Note: If the file does not download, see if you have an ad popup blocker running. If so, disable it, and download the file again.

Input Files

This page displays all the parameters and files used.

DNA and DNA + Protein

• Panel Files
• FASTQ Files
• Lane Assignments

Merge Samples

• Input Runs