tb_day3_control_rep1
Poly(A) RNA was extracted using magnetic oligo-dT beads and was fragmented by heating. Fragmented RNA was used for first-strand cDNA synthesis using random primers, followed by second-strand cDNA synthesis to generate double-stranded cDNA (dscDNA), where the second strand was marked with deoxyuridine triphosphate (dUTP). The resulting dscDNA was subjected to bead-based cleanup and A-tailing. Unique dsDNA adapters were ligated to the library fragments, followed by bead-based cleanup. The adapter-ligated library was amplified through polymerase chain reaction (PCR) and purified using a bead-based cleanup. Each library was quantified using qPCR and pooled with the other samples. The pooled library was single-end sequenced using Illumina NextSeq (Illumina Inc., San Diego, CA, USA).