TY - JOUR A1 - Berdoz, J A1 - Monath, T P A1 - Kraehenbuhl, J P T1 - Specific amplification by PCR of rearranged genomic variable regions of immunoglobulin genes from mouse hybridoma cells. Y1 - 1995/04/01 JF - Genome Research JO - Genome Research SP - 256 EP - 264 VL - 4 IS - 5 UR - http://genome.cshlp.org/content/4/5/256.abstract N2 - We have designed a novel strategy for the isolation of the rearranged genomic fragments encoding the L-VH-D-JH and L-V kappa/lambda-J kappa/lambda regions of mouse immunoglobulin genes. This strategy is based on the PCR amplification of genomic DNA from mouse hybridomas using multiple specific primers chosen in the 5'-untranslated region and in the intron downstream of the rearranged JH/J kappa/lambda sequences. Variable regions with intact coding sequences, including full-length leader peptides (L) can be obtained without previous DNA sequencing. Our strategy is based on a genomic template that produces fragments that do not need to be adapted for recombinant antibody expression, thus facilitating the generation of chimeric and isotype-switched immunoglobulins. ER -