GSM5412360: Nsp7_rep_1; Homo sapiens; RNA-Seq
Total RNA was extracted using Trizol reagent (Sigma) and treated with DNase I (NEB) following the manufacturer's protocol. For 3' RNA sequencing libraries, 75ng of RNA were used as input  following CelSeq2(Hashimshony et al., 2016) protocol, changing the UMI to 6 bases. Sequencing was performed on Illumina NextSeq 500. For total RNA libraries 40ng of rRNA depleted RNA was used as a starting material for library preparation. NEBNext rRNA Depletion kit( NEB #E7405) was used to deplete ribosomal RNA. Libraries were prepared using the NEXTFLEX® Rapid Directional RNA-Seq Kit 2.0 by PerkinElmer, following the manufacturer's instructions. The samples were sequenced by Novogene (Novogene Corporation Inc. 8801 Folsom BLVD, Suite 290, Sacramento, CA 95826) with Hiseq-4000. 3' RNA sequencing libraries stranded, CelSeq2 based. For total RNA libraries, stranded  ligation-based.