Amplicon-seq of human ocular adnexal marginal zone lymphoma of MALT subtype
For multiplex PCR two customized Ion AmpliSeq primer pools for a total of 1255 amplicons were used (Life Technologies, Darmstadt, Germany). Primer sequences are available from the authors upon request. The DNA input per reaction was 10 ng. Multiplex PCR was performed (23 cycles) for 24 target genes (Table 2), most of which are involved in NF-kB signaling. Libraries were prepared with the NEBNext Ultra DNA Library kit (NEB, Ipswich, UK) for sequencing on the Illumina platform. Purified libraries were quantified with the KAPA library quantification kit (Peqlab, Erlangen, Germany). Library size and quality was determined with the Agilent Bioanalyzer high sensitivity DNA assay (Agilent, Santa Clara, USA). Sequencing was performed on an Illumina MiSeq apparatus with 150 bp paired end reads.