RNA-seq Illumina prepared with PIP-seq, hashtagged fixation protocol vs fresh panel, of homo sapiens: PBMCs
Following WTA cDNA of PIP-seq captures, the supernatant of first SPRI cleanup which contained Hashtag cDNA was further processed as per manufacturers instructions. Briefly, following another round of SPRI purification, libraries were amplified (8 cycles) using primers that appended illumina adapters. Libraries were SPRI cleaned, evaluated using high-sensitivity D1000 ScreenTape on 4150 Tapestation (Agilent Technologies) and sequenced either on Illumina NovaSeq SP (54-bp Read 1, 8-bp i7 index, 8-bp i5 index, and 67-bp Read 2) or NovaSeq 2000 (28-bp Read 1, 10-bp i7 index, 10-bp i5 index, and 90-bp Read 2).