title: R_KO_1_3
description: Protocols: Cells were plated on 10 cm plates and grown for 48 hours. Subsequently, the cells were pelleted, washed twice with PBS, and stored at -80°C. Total RNA was isolated using the RNeasy Mini Kit (QIAGEN) according to the manufacturer's instructions.The concentration of isolated RNA was measured by NanoDrop. This step was performed by Novogene Europe. Messenger RNA was purified from total RNA using poly-T oligo-attachedmagnetic beads. After fragmentation, the first strand cDNA was synthesizedusing random hexamer primers, followed by the second strand cDNAsynthesis using either dUTP for directional library or dTTP for non-directional library.For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification. The library was checked with Qubit and real-time PCR for quantificationand bioanalyzer for size distribution detection. Quantified libraries were and sequenced on Illumina platforms, according to effective library concentration and data amount.
attributes.tag: replicate
attributes.value: biological replicate 3
attributes.tag: disease
attributes.value: normal
attributes.tag: organism
attributes.value: Homo sapiens
attributes.tag: organism
attributes.value: Homo sapiens
attributes.tag: collection date
attributes.value: not collected
attributes.tag: scientific_name
attributes.value: Homo sapiens
attributes.tag: cell type
attributes.value: embryonic kidney
attributes.tag: common name
attributes.value: human
attributes.tag: organism part
attributes.value: kidney
attributes.tag: developmental stage
attributes.value: embryo stage
attributes.tag: genotype
attributes.value: female, immortalised by adenovirus 5 integration
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: genetic modification
attributes.value: LYAR knockout - rescued
attributes.tag: cell line
attributes.value: HEK293
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected