Name: Sample 24_p
Instrument: Illumina NovaSeq 6000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: PolyA
Layout: PAIRED
Construction protocol: All fibroblast derived cell lines were treated with different stimuli for 4 hours. Cells were washed with PBS and then lysed in RNA Lysis Buffer (Zymo Research). Cells were stimulated with 1 ug/mL of High-Molecular Weight poly(I:C) (InvivoGen, Cat. Code: tlrl-pic), transfected with 2 uL/mL Lipofectamin 2000 (Thermo Fisher Scientific), for 4 hours before collection. Total RNA was extracted using the Quick-RNA MicroPrep kit (Zymo Research, Cat. No. R1051). Libraries were produced using the Kapa Stranded mRNA-seq Kit (24RXN) (KK8420 ROCHE-07962193001) with the KAPA Unique Dual-Indexed Adapter Plate (KK8726-08861862001) and KAPA Adapter Dilution Buffer (KK8721-08278539001), starting with 500 ng of total RNA (samples #1-4 were started with 275-337 ng total). For all samples, except sample #39, fragmentation was performed as recommended for 7 minutes. Fragmentation for sample #39 was set to 5 minutes (RIN 6.0). UDI adapters were diluted to 350nM as instructed in the protocol. Final cDNA amplification was performed with 13 PCR cycles. Libraries were normalized and pooled at 10 nM.