RNA-seq Illumina prepared with PIP-Seq of homo sapiens: PBMCs
Libraries were prepared for Illumina sequencing as per the manufacturers instructions (PIPseq T20 Single Cell RNA kit v4.0). Briefly, 30ng of PBMC cDNA and 100 ng of K562:3T3 cDNA were input into the PIP-seq library prep module to undergo fragmentation, end repair, A-tailing, and adapter ligation and underwent indexing PCR using 11 cycles for PMBC and 9 cycles for K562:3T3. Final libraries were quantified using Qubit and fragment sizes were verified using a high-sensitivity D1000 ScreenTape on 4150 Tapestation (Agilent Technologies) prior to equimolar pooling and sequencing at BCL on an either an Illumina NovaSeq X (28-bp Read 1, 10-bp i7 index, 10-bp i5 index, and 90-bp Read 2), 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).