title: Gene-specific PLCG2 targeted RNAseq of human blood
design: Peripheral blood mononuclear cells (PBMC) were isolated from participants by gradient centrifugation. RNeasy kits (Qiagen) were used to isolate RNA. Full-length PLCG2 cDNA was synthesized by reverse transcription with SuperScript III Reverse Transcriptase (Thermo Fisher) and a PLCG2-specific primer that recognizes a region of the 3' untranslated region that is conserved in almost all PLCG2 transcripts. Full-length PLCG2 was amplified from cDNA by long-range PCR with TaKaRa LA Taq (Takara Bio) and custom primers.  Sequencing libraries were generated using Nextera library prep kits (Illumina) and were sequenced with 150bp paired end reads on a MiSeq instrument (Illumina). Reads were aligned to the PLCG2 cDNA sequence (hg19) using STAR and parsed for alternate transcripts containing deletions.
identifiers.namespace: NIAID-phs003807
identifiers.id: Proband7