Name: GSM7473115
Instrument: NextSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: RNA extraction from collected cells of wild-type and Cgsnf2Δ strains was done using Qiagen RNeasy kit following manufacturers' protocol. The NEBNext Poly(A) mRNA Magnetic Isolation Module is used for the enrichment of mRNA. Poly (A) purified RNA sample is used for library preparation using NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® as per manufacturer protocol. In short, following purification, the RNA is fragmented using divalent cations under elevated temperature. Next, the cDNA is synthesized using Reverse transcriptase and random hexamers in a first-strand synthesis reaction. Subsequently, the cDNA is converted to double-stranded cDNA where Uracil is added instead of Thymine. The strand specificity is preserved by a USER enzyme-based digestion of the second strand, thereby leaving one functional strand which maps to the DNA strand from which it was transcribed from. The USER digested single strand molecules were enriched and indexed in a limited cycle PCR, followed by AMPure bead purification to create the final cDNA library for sequencing.