title: GSM4827296: HK2 ANXA2 knockdown rep1; Homo sapiens; RNA-Seq
library_construction_protocol: HK2 cells were ground into fine powder before RNA extraction. Total RNA was extracted using TRIzol (Ambion)（Invitrogen, Carlsbad, CA, USA）. The RNA was further purified with two phenol-chloroform treatments and then treated with RQ1 DNase (Promega, Madison, WI, USA) to remove DNA. The purified RNAs were then iron-fragmented at 95 ˚C followed by end repair and 5' adaptor ligation. Then, reverse transcription was performed with RT primer harboring 3' adaptor sequence and randomized hexamer. The cDNAs were purified, amplified, and stored at −80 ˚C until required for sequencing. For each sample, 1 μg of the total RNA was used for RNA-sequencing (RNA-seq) library preparation. Polyadenylated RNAs were purified and concentrated using oligo(dT)-conjugated magnetic beads (Invitrogen, Carlsbad, CA, USA) before directional RNA-seq library preparation.