GSM3554279: HSV-1_1H_RNASeq_rep2; Human alphaherpesvirus 1; RNA-Seq
Total RNA was extracted by using EASYspin RNA Mini Kit (Aidlab RN07). Briefly, tissue was triturated by trituration equipment for 20 s in lysis buffer provided by kit and then centrifuged at 12000 rpm, 4℃ for 5 min. Liquid between precipitates on the bottom and oil on the top was taken out and pipetted 10 times using 1 mL syringe. Entire volume of liquid was added into adsorption column and total RNA eluted with 50 μL RNase-free water. RNA-seq libraries were constructed by NEBNext Poly (A) mRNA Magnetic Isolation Module (NEB E7490) and NEBNext Ultra II Non-Directional RNA Second Strand Synthesis Module (NEB E6111). Briefly, poly-A mRNA was prepared with poly-T magnetic beads and transformed into cDNA with first and second strand synthesis. Newly synthesized cDNA were purified by AMPure XP beads (1:1) and eluted in 50 μL nucleotide-free water. RNA-seq libraries were the same as ChIP-seq library construction described previously except the sequencing depth of 20 million reads per library. RNA-seq libraries were sequenced by Illumina Hiseq X Ten platform with pair-end reads of 150 bp.
GEO Accession
GSM3554279