title: GSM8282326: JUP knockdown1; Homo sapiens; RNA-Seq
library_name: GSM8282326
library_construction_protocol: SAECs after knockdown were washed with PBS and total RNA from the samples wereextracted using the RNeasy Mini Kit (Qiagen) according to the manufacturer's instructions. The RNA concentrations and purity were measured on a NanoDrop ND-1000 spectrophotometer, followed by tapestation RNA quality was determined initially using nanodrop, followed by Agilent 2200 and TapeStation Analysis Software 3.2. All samples had excellent quality with RNA integrity number (RIN) 10. Libraries for RNA-Seq were prepared with KAPA mRNA-Seq Hyper Prep Kit according to manufacturer's instructions.  cDNA generation is followed by end repair to generate blunt ends, A-tailing, adaptor ligation and PCR amplification. Different adaptors were used for multiplexing samples in one lane. Libraries for RNA-Seq were prepared with KAPA mRNA-Seq Hyper Prep Kit. The workflow consists of (1) mRNA enrichment and fragmentation, (2) first strand cDNA synthesis using random priming followed by second strand synthesis converting cDNA:RNA hybrid to double-stranded cDNA (dscDNA), and (3) incorporating dUTP into the second cDNA strand. cDNA generation is followed by end repair to generate blunt ends, A-tailing, adaptor ligation and PCR amplification.