作者: Hamilton, A. ; Adair, J. ; Weir, N. ; Moss, M. ; Tempest, P. ; Fernie, M. ; Forster, S. ; Armour, K. ; Steven, J. ; King, S. ; Carr, F. ; Wallace, T. P. ; Harris, W. ; King, D. ; White, P. ; Mcgregor, D. ; Turner, A.
The basic steps for isolation of antibody variable region cDNAs from a murine hybridoma are described.Protocols are given for resurrecting mammalian cells from liquid nitrogen, cell culture of hybridoma cells, preparation of RNA from hybridoma cells, preparation of 1st-strand primer-directed cDNA and antibody variable region amplification, and cloning antibody variable region sequences.These protocols can be easily adapted for human antibody variable region cloning.