VDJ CDR3 Protocols

For Single Cell Preparation and Preservation Protocols, see Sample Prep and Preservation Section

For Cell Labelling Protocols, see BD® AbSeq Intracellular and Cell Surface Protein Profiling Section or the BD® Single-Cell Multiplexing Kit (SMK and Flex SMK) section

Cell Capture

Perform cell capture on the single lane cartridge using :

•  With scanner : BD Rhapsody™ Single-Cell Analysis System Instrument User Guide (Doc ID: 23-21336) STOP after the section “Washing the BD Rhapsody™ Enhanced Cell Capture Beads” and follow the protocols below under Sequencing Library Preparation

Or,

• Without scanner : BD Rhapsody™ Express Single-Cell Analysis System Instrument User Guide (Doc ID: 23-21332) STOP after the section “Washing the BD Rhapsody™ Enhanced Cell Capture Beads” and follow the protocols below under Sequencing Library Preparation 

If using a 8-lane High Throughput cartridge, perform cell capture using : 

• Without scanner : BD Rhapsody™ HT Xpress System Instrument User Guide for Scanner-free workflow(Doc ID : 23-24256) STOP after the section “Washing the BD Rhapsody™ Enhanced Cell Capture Beads” and follow the protocols below under Sequencing Library Preparation

Or,

• With scanner : BD Rhapsody™ HT Single-Cell Analysis System Instrument User Guide(Doc ID: 23-24257) STOP after the section “Washing the BD Rhapsody™ Enhanced Cell Capture Beads” and follow the protocols below under Sequencing Library Preparation

Sequencing Library Preparation

For use with v1 beads only, do not use with enhanced beads

Interested in VDJ CDR3 with enhanced beads? Contact us at scomix@bd.com

IMPORTANT: The protocol described herein has not been thoroughly tested or optimized and is provided as is and without warranty. Any use of this protocol is at the risk of the user. BD Biosciences reserves the right to change specifications at any time. Information in the protocol is subject to change without notice. BD Biosciences assumes no responsibility for any errors or omissions. In no event shall BD Biosciences be liable for any damages in connection with or arising from the use of this protocol.

1. VDJ CDR3 Library Preparation Protocol  (Human)
   • This protocol provides instructions on creating a single-cell mRNA library with the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina® sequencers. It is also intended to provide a method to enable a high-throughput detection of VDJ recombination events by generating sequencer-ready libraries specific to the CDR3-specific antigen binding regions of B and T cells. 
   • Doc ID: 23-22202-00
   • Revision date: 01/2020
2. VDJ CDR3 and BD AbSeq Library Preparation Protocol  (Human)
   • This protocol provides instructions on creating single-cell mRNA and BD Abseq libraries with the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina® sequencers. It is also intended to provide a method to enable a high-throughput detection of VDJ recombination events by generating sequencer-ready libraries specific to the CDR3-specific antigen binding regions of B and T cells. 
   • Doc ID: 23-22203-01
   • Revision date: 05/2020
3. VDJ CDR3 and Sample Tag Library Preparation Protocol  (Human)
   • This protocol provides instructions on creating single-cell mRNA and Sample Tag libraries with the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina® sequencers. It is also intended to provide a method to enable a high-throughput detection of VDJ recombination events by generating sequencer-ready libraries specific to the CDR3-specific antigen binding regions of B and T cells. 
   • Doc ID: 23-22205-00
   • Revision date: 01/2020
4. VDJ CDR3, Sample Tag, and BD AbSeq Library Preparation Protocol (Human)
   • This protocol provides instructions on creating single-cell mRNA, Sample Tag, and BD Abseq libraries with the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina® sequencers. It is also intended to provide a method to enable a high-throughput detection of VDJ recombination events by generating sequencer-ready libraries specific to the CDR3-specific antigen binding regions of B and T cells for human samples. 
   • Doc ID: 23-22204-01
   • Revision date: 05/2020
5. VDJ CDR3 Library Preparation Protocol  (Mouse)
   • This protocol provides instructions on creating a single-cell mRNA library with the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina® sequencers. It is also intended to provide a method to enable a high-throughput detection of VDJ recombination events by generating sequencer-ready libraries specific to the CDR3-specific antigen binding regions of B and T cells for mouse samples.
   • Doc ID: 23-22687-00
   • Revision date: 05/2020
6. VDJ CDR3 and Sample Tag Library Preparation Protocol  (Mouse)
   • This protocol provides instructions on creating single-cell mRNA and Sample Tag libraries with the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina® sequencers. It is also intended to provide a method to enable a high-throughput detection of VDJ recombination events by generating sequencer-ready libraries specific to the CDR3-specific antigen binding regions of B and T cells for mouse samples. 
   • Doc ID: 23-22688-00
   • Revision date: 05/2020

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