454 GS Junior sequencing
Analysing the BR repertoire in HD of different ages
DNA from sorted naive B cells (CD19+ CD27-) from periperhal blood was amplified using forward VH1-6 FR1 (BIOMED-2) primers and the JH consensus reverse primer. For each healthy donor 3-6 independent PCR reaction were performed. Samples were purified by gel extraction and AMPure XP beads and sequenced using 454 GS junior sequencing with the Lib-A kit.  The deposited data are untrimmed. In the analysis of IJspeert et al Front Immunol 2015 and IJspeert et al Front. Immunol. 7:410. doi: 10.3389/fimmu.2016.00410 are deposited. samples were demultiplexed and filtered using  CLC genomic workbench. Low quality reads (limit =0.05), reads with ambigious nucleotides (max 2), and sequences shorter than 100nt or longer than 800nt were removed. In addition the adaptor sequences were removed and sequences were trimmed 20-25 nt.