title: NextSeq 550 paired end sequencing; RNA-seq of Zbtb10 Knockdown and Control cDC1 dendritic cells (Cd8+)
design: RNA-seq of Zbtb10 Knockdown and Control cDC1 dendritic cells (Cd8+)
library_construction_protocol: The RNA-seq library preparation was performed for Zbtb10 KD and control cDC1 MutuDCs at 2h and 6h after CpG activation. As we did time kinetics, we included three independent biological replicates to identify the Zbtb10 depletion mediated global transcriptome changes.  Cells were lysed using Lysis buffer (LBP, Takara) Followed by removal of genomic DNA, using gDNA eliminator column, the RNA was then ethanol precipitated in a nucleospin column (Takara). The total RNA was eluted in RNase free water and checked for concentration and purity using Nanodrop. For RNA-seq library preparation 2 Âµg of total RNA was used to isolate mRNA through magnetic beads using mRNA isolation kit (PolyA mRNA isolation Module, NEB) followed by RNA-seq library preparation using mRNA library preparation kit (NEB) strictly following the vendor recommended protocol.