GSM4495376: KO_EGF_1; Mus musculus; RNA-Seq
Organoids were dissociated using Tryp-LE and RNA was extracted using the Qiagen Rneasy kit (cat# 74104) Organoids were dissociated to single cells using TrypLE Select (Life Technologies 12563011). Organoids were incubated with TrypLE for 5 minutes at 37°C, and washed in PBS. RNA was isolated from organoids using the RNeasy Mini Kit (Qiagen 74104). RNA purity and quantitation was assessed using the Nanodrop ND-800 (Nanodrp Technologies, Inc, Wilmington, D) and RNA integrity was assessed using the 5200 Fragment Analyzer System and the RNA high-sensitivity chip (Agilent, Santa Clara, CA, USA). Sequencing libraries were generated using 100 ng of total RNA and the Universal RNA-Seq library preparation Kit with multiplexing primers, according the manufacturer's protocol (Nugen technologies, Inc, San Carlos, CA, USA). Library size distribution and concentration were assessed using the Fragment Analyzer System and the DNA high-sensitivity chip. Libraries were multiplexed at a density of 8 per flow-cell lane and sequenced on the HiSeq 4000 to generate single end 50bp reads per the manufacturer's instructions (Illumina, San Diego, CA, USA).
GEO
GSM4495376
GEO Accession
GSM4495376