Cloning and expression of murine Ig genes from single B cells

T Tiller, CE Busse, H Wardemann - Journal of immunological methods, 2009 - Elsevier
We have established a highly efficient 96-well format based strategy to characterize the
expressed murine antibody repertoire by combining immunoglobulin (Ig) gene cloning with
antibody expression and reactivity profiling at the single cell level. Individual mouse B
lineage cells are isolated based on defined surface marker expression patterns by
fluorescence-activated cell sorting (FACS) and corresponding full-length Ig heavy (H) and Ig
light (L) chain variable (V) region gene transcripts are amplified by RT-PCR. Cloning of the …