Box 1: Kit Components
| Component | Description | Cap Color | Conc | Total Volume |
|---|---|---|---|---|
| Forward GSP BCR-Mark30 primer mix (human) | Mix of forward gene-specific primers to amplify CDR1-2-3 region of IGH, IGK, and IGL and 100-250 bp ORF region of 30 B cell marker genes | n/a | 20X | 30 µl |
| scAIR BCR Positive Control RNA | RNA from B (CD19+) cells purified from normal donor PBMC | n/a | 20 pg/μl | 200 μl |
| SPRI bead Dilution Buffer | Buffer to dilute SPRIselect beads four-fold prior to use | n/a | 1X | 10 ml |
| Water | Milli-Q purified water to prepare reaction mixes | White | NA | 10 ml |
| dNTP mix | Mix of dATP, dGTP, dCTP and dTTP, 10mM each | White | 20X to 50X | 50 µl |
| RT-EXT Buffer | cDNA synthesis and Forward gene-specific primer extension reaction buffer | Green | 5X | 250 µl |
| Reverse Transcriptase | Multiplex-optimized M-MLV Reverse Transcriptase (RT) | Green | 20X | 20 µl |
| Hot-Start RT Aptamer | Aptamer which inactivates RT at room temperature but loses inhibition activity at 50°C | Green | 20X | 20 µl |
| PCR Buffer | Reaction Buffer for 1st and 2nd PCR steps | Red | 5X | 350 µl |
| DNA polymerase | Multiplex-optimized thermostable hot-start DNA polymerase | Red | 100X | 25 µl |
| Anchor Primer Mix | Forward and Reverse Anchor-specific primers (AP1 and AP2) for 1st PCR | Red | 10X | 250 µl |
| Primer Removal Enzyme | Enzyme mix for primer removal | Orange | 20X to 50X | 500 µl |
| Primer Removal Buffer | Buffer to prepare Primer removal master mix with Primer Removal Enzyme | Orange | 10X | 1 ml |
| Forward SeqDNA NGS Primer | Forward SeqDNA NGS Primer for cDNA sequencing | Blue | 100 µM | 50 µl |
| Reverse SeqDNA NGS Primer | Reverse SeqDNA NGS Primer for cDNA sequencing | Blue | 100 µM | 50 µl |
| Forward SeqIND NGS Primer | Forward SeqIND NGS Primer for cDNA sequencing | Blue | 100 µM | 50 µl |
| Reverse SeqIND NGS Primer | Reverse SeqIND NGS Primer for cDNA sequencing | Blue | 100 µM | 50 µl |
Box 2: Kit Components
| Component | Description | Conc | Total Volume |
|---|---|---|---|
| scAIR-BCR-Mark30 Rev GSP Master Plate | 96-well plate, each well contains 50 µl of hybridization buffer and a mix of barcoded (well-specific and VBC barcodes) reverse gene-specific primers (Rev GSPs) designed for the C region of IGH, IGK, and IGL, and the ORF region of 30 B cell marker mRNAs | 1X | 50 µl/well in a 96-well plate |
| Dual Index Primer Set Plate | 96 Dual DNA/RNA UD Indexes primer pairs, pre-aliquoted and dried in a 96-well plate | NA | One 96-well plate (dried) |
Kit contents should be stored at -20°C. Shelf life is 1 year from the date of receipt.
Last modified:
14 May 2025
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