RNA-seq of Prinia subflava: shell gland tissue
Poly (A)-containing mRNA molecules were purified from total RNA using poly (T) oligo-attached magnetic beads, after which mRNAs were fragmented into small pieces using a fragmentation reagent. First strand cDNA was synthesized using reverse transcriptase and random hexamer primers followed by second strand synthesis, end repair, and single 3 adenylation of blunt ends. Indexed adapters were then ligated followed by a PCR reaction to enriches for fragments with adapters ligated to both ends. Fragment libraries were checked and quantified using an Agilent Technologies 2100 Bioanalyzer prior to pooling of individual sample libraries with different barcodes/indexes for sequencing in one lane.