title: GSM4290756: S4C_9_S9; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from ~5.00x10^6 cells using the Quick-RNA Miniprep Kit (Zymo R1055) according to manufacturer's instructions. RNA quality was determined with Agilent Tapestation ScreenTape (Agilent 5067-5576). 1µg of total RNA with ERCC Spike-ins (ThermoFisher 4456740) was used to prepare libraries with KAPA stranded RNA-seq kit with RiboErase (HMR) (KAPA Biosystems KK8483) according to manufacturer's instructions. Briefly, total RNA was hybridized and rRNA depleted followed by DNase digestion. RNA is then fragmented to a size of ~100-200bp and primed for first strand synthesis. First and second strand synthesis is performed and the reaction is incubated overnight in A-tailing buffer at 4°C. The next day, A-tailing is performed followed by adapter ligation (NEB E735L) and library amplification for 12 cycles. After final cleanup, libraries are run on Agilent Tapestation DNA D1000 ScreenTape (Agilent 5067-5582) to confirm fragment size and quantified using a Qubit 2.0 fluorometer. Libraries are submitted to the UTSWMC McDermott Center Sequencing Core and 75bp pair-end reads were generated for both libraries.
attributes.tag: GEO Accession
attributes.value: GSM4290756