GSM7616058: NP-specific Germinal Center B cell, Day12 Immunized control,rep5; Mus musculus; RNA-Seq
Cells were sorted into 12.5 ul of lysis buffer and cDNA was generated using the Takara Smart-Seq HT kit (Takara: 634438) following manufactures instructions, with 15 cycles of cDNA amplification. Smart-Seq cDNA was assessed for quality on an Agilent Fragment Analyzer 5300 using the High sensitivity NGS kit (Agilent: DNF-474-1000). Samples that passed QC with full length cDNA proceeded to library preparation using Illumina Nextera DNA prep (Illumina: 20018705) with UDI indexes (Illumina: 20042666) added using 8 PCR cycles. Libraries were pooled and sequenced on an Illumina NextSeq 2000 using a P3 100 cycle flow cell (Illumina: 20046812), 2x59 bp, for an average of ~35 million reads per sample. Libraries were demultiplexed using the NextSeq 2000 Dragen server and BCL Convert (v 3.8.4).