Product Description
This kit generates full-length cDNA from 1–500 rabbit B cells—or 10 pg–5 ng total RNA—in a single reaction tube. It then specifically amplifies the B cell receptor IgG heavy- and κ light-chain variable regions and, without purification, produces transfection-ready DNA fragments suitable for direct transient expression in mammalian cells.
Applications
Recombinant expression and validation of B cell receptors (BCRs)
Product Advantages
- Ultra-low input: Requires only 1–500 rabbit B cells or 10 pg–5 ng total RNA as starting material.
- High pairing efficiency: Heavy and light chains co-amplified with > 90 % pairing success.
- Purification-free workflow: Generates transfection-ready DNA cassettes without any cleanup steps.
Kit Composition & Storage Conditions (96 Reactions)
| Component | Volume | Storage |
|---|
| dNTP Mix | 192μL | -20℃ |
| Cell Lysis Buffer | 96μL | -20℃ |
| RT Primer | 38.4μL | -20℃ |
| DTT | 96μL | -20℃ |
| Murine RNase Inhibitor | 38.4μL | -20℃ |
| Reverse Transcriptase | 38.4μL | -20℃ |
| 2 × PCR Mix | 10mL | -20℃ |
| PCR1 Primer Mix | 480μL | -20℃ |
| PCR2 rHeavy Primer Mix / PCR2 rLight Primer Mix | 96μL each | -20℃ |
| PCR3 rHeavy TAD Mix / PCR3 rLight TAD Mix | 192μL each | -20℃ |
| PCR3 TAD Primer Mix | 192μL | -20℃ |
| RNase-Free water | 6mL | -20℃ |
| TS Primer | 96μL | -80℃ |
| 5 × RT Buffer | 384μL | -80℃ |
User‑Supplied Materials
- Sample: Cell sample or equivalent micro‑quantity RNA
- Equipments: shaker, mini‑centrifuge, thermal cycler, etc.
