title: GSM9604045: rhozero-7-2-24hpi-Exp161; Homo sapiens; RNA-Seq
library_construction_protocol: RNA extractions were later performed under RNAse-free conditions (Thermo Fisher Scientific; 7002) and RNA eluted in 30µL. Performed rRNA depletion was performed using QIAGEN FastSelect rRNA HMR Kit (Qiagen; 334386). RNA sequencing libraries were constructed with the NEBNext Ultra II RNA Library Preparation Kit for Illumina (New England Biolabs; E7770L).  Enriched RNAs are fragmented for 15 min at 94 °C. First strand and second strand cDNAs were subsequently synthesized. cDNA fragments were end repaired and adenylated at 3'ends, and universal adapters are ligated to cDNA fragments, followed by index addition and library enrichment with limited cycle PCR. Sequencing libraries were validated using the Agilent Tapestation 4200 (Agilent Technologies, Palo Alto, CA, USA), and quantified using Qubit 2.0 Fluorometer (Thermo Fisher Scientific, Waltham, MA, USA) as well as by quantitative PCR (KAPA Biosystems, Wilmington, MA, USA). The sequencing libraries were multiplexed and clustered on the flowcell. After clustering, the flowcell was loaded on the Illumina NovaSeq instrument and the samples were sequenced using a 2x150 Paired-End (PE) configuration.