WTA Protocols

Cell Preparation and Cell Labeling

1. Preparing Single Cell Suspensions

• This protocol describes preparation of single cell suspensions of commonly used immune cell types. A single cell suspension can be loaded into a BD Rhapsody™ Cartridge. This protocol does not describe tissue dissociation.
• Doc ID: 210964 Rev. 1.0
• Revision date: 07/2018

1. Single Cell Labelling with the BD Single-Cell Multiplexing Kit

• The BD Single-Cell Multiplexing Kit (Cat. No. 633781) utilizes an innovative antibody-oligo technology to provide higher sample throughput for single cell library preparation. Every antibody-oligo in the kit, referred to as a Sample Tag, has a unique sample barcode conjugated to a human universal antibody. Up to 12 samples can be labelled and pooled prior to single cell capture with the BD Rhapsody™ Single-Cell Analysis system or other single cell analysis systems.
• Doc ID: 210970 Rev. 2.0
• Revision date: 02/2019

1. Single Cell Labelling with BD™ AbSeq Ab-Oligos

• This protocol describes use of BD AbSeq Ab-Oligos (antibody-oligonucleotides).
• Doc ID: 214394 Rev. 1.0
• Revision date: 07/2018

1. Single Cell Labelling with the BD Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos (1 plex to 40 plex)

• This protocol describes use of BD AbSeq Ab-Oligos (antibody-oligonucleotides)(1 plex to 40 plex) with the BD® Single-Cell Multiplexing Kit.
• Doc ID: 214419 Rev. 2.0
• Revision date: 02/2019

1. Single-Cell Labeling with BD Single-Cell Multiplexing Kit and BD® AbSeq Ab-Oligos (41 plex to 100 plex)

• This protocol describes the use of BD AbSeq Ab-Oligos (antibody-oligonucleotides)(41 plex to 100 plex) with the BD® Human Single-Cell Multiplexing Kit.
• Doc ID: 23-22354-00
• Revision date: 05/2020

cDNA Synthesis

1. Single Cell Capture and cDNA Synthesis with the BD Rhapsody Express Single-Cell Analysis System

• This protocol describes cell loading in the BD Rhapsody™ Cartridge and single cell capture with the BD Rhapsody™ Express Single-Cell Analysis system.
• Doc ID: 210967 Rev. 1.0
• Revision date: 07/2018

1. Single Cell Capture and cDNA Synthesis with the BD Rhapsody Single-Cell Analysis System

• This protocol describes cell loading in the BD Rhapsody™ Cartridge and single cell capture with the BD Rhapsody™ Single-Cell Analysis system with the BD Rhapsody™ Scanner.
• Doc ID: 210966 Rev. 1.0
• Revision date: 07/2018

Sequencing Library Preparation

1. mRNA Whole Transcriptome Analysis (WTA) Library Preparation Protocol

• This protocol provides instructions on creating a single cell whole transcriptome mRNA library after cell capture on the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina sequencers. 
• Doc ID: 23-21711-00
• Revision date: 07/2019

1. mRNA Whole Transcriptome Analysis (WTA) and AbSeq Library Preparation Protocol

• This protocol provides instructions on creating single cell whole transcriptome mRNA and AbSeq protein detection libraries after cell capture on the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina sequencers. 
• Doc ID: 23-21751-00
• Revision date: 07/2019

1. mRNA Whole Transcriptome Analysis (WTA) and Sample Tag Library Preparation Protocol

• This protocol provides instructions on creating single cell whole transcriptome mRNA and Sample Tag libraries after cell capture on the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina sequencers. 
• Doc ID: 23-21712-00
• Revision date: 07/2019

1. mRNA Whole Transcriptome Analysis (WTA), AbSeq, and Sample Tag Library Preparation Protocol

• This protocol provides instructions on creating single cell whole transcriptome mRNA, AbSeq, and Sample Tag libraries after cell capture on the BD Rhapsody™ Single-Cell Analysis System or the BD Rhapsody™ Express Single-Cell Analysis System for sequencing on Illumina sequencers. 
• Doc ID: 23-21752-00
• Revision date: 07/2019