Ig-Seq (IgG) of mus musculus (BALB/c): juvenile female spleen
Targeted Sequencing of IgG1, IgG2a, IgG2b, IgG2c and IgG3. Libraries are prepared via RT-PCR. First-strand cDNA synthesis was performed with isotype-specific primers introduced in the RT step. Sequence amplification was conducted via a nested PCR approach: First, sequences were amplifiied using a multiplexed foward primer set against the framework 1 region and one specific reverse primer. In a second PCR step, Illumina's sequencing adapters are completed by targeting 3' and 5' constant overhang regions introduced by the primers of the previous PCR step.