A simple way to increase recovery of the expressed VH and VL genes in single-sorted human B cells
| Autor: | Sergey V Guselnikov, Tatyana N Belovezhets, Sergey V Kulemzin, Andrey A Gorchakov, Alexander V Taranin |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 67, Iss 4, Pp 184-187 (2019) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/btn-2019-0079 |
| Popis: | Cloning VH and VL genes from individual antigen-specific B cells is an attractive approach for producing monoclonal antibodies of the desired specificity. Current RT-PCR protocols, however, result in the successful identification of VH and VL gene pairs in about half of the sorted cells. Here, we demonstrate that single-cell RT-PCR is likely affected by stochastic factors, and that running PCRs in triplicate results in successful amplification of the expressed VH and VL genes in 90–100% of single sorted human B cells. |
| Databáze: | Directory of Open Access Journals |
| Externí odkaz: |
|