title: PBMC heavy chain antibody repertoire sequencing of llama glama
design: Llama PBMCs were isolated using Ficoll-Paque gradient and RNA was extracted using an Qiagen RNA extraction kit following manufacturer's protocol. RNA was reverse-transcribed using a custom race adapter, which contains UMIs and deoxyUracil modifications. The obtained cDNA was digested with Uracil DNA glycosylase for 40min at 37C and purified. cDNA, roughly estimated to stem from about 20000 B-cells, was used as input for a PCR amplification. Sequencing libraries were generated using the NEBNext Ultra DNA Library Prep Kit. The obtained sequence libraries were purified using an SDS page gel and a gel extraction kit. The libraries were pooled and sequenced on an Illumina Miseq sequencer using asymmetric long read option (400 x 100bp).