title: GSM8416755: HK2-OEHNRNPF-1; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted according to the Trizol method, and 1 μL of total RNA was used as the template, reverse transcribed to cDNA, and continued to be amplified by using cDNA as the template, sequentially 95 ℃ for 3 min, 1 cycle, 95 ℃ for 10 s, and 62 ℃ for 40 s, for a total of 40 cycles. The mRNA levels of each group were calculated using 2-ΔΔCt method with β-actin as internal reference. Library preparation, sequencing information (eg. cDNA libraries were prepared using the SMARTer Stranded Total RNA-Seq Kit version 2 (634413; Takara/Clontech) according to the manufacturer's procedure. 150 nt double-end sequencing was performed on the Illumina NovaSeq system) 1 μg of total RNA was used for RNA-seq library preparation. mRNAs were captured by VAHTS mRNA capture Beads (Vazyme, N401).The purified RNA was treated with RQ1 DNase (Promega) to remove DNA before used for directional VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina （NR605） Polyadenylated mRNAs were purified and fragmented. Fragmented mRNAs were converted into double strand cDNA. Following end repair and A tailing, the DNAs were ligated to Adaptor (N323). After purification of ligation product and size fractioning to 300-500bps, the ligated products were amplified and purified, then quantified and stored at -80℃ before sequencing. The strand marked with dUTP (the 2nd cDNA strand) is not amplified, allowing strand-specific sequencing