BCK_3
RNA library for metatranscriptome sequencing was prepared as rRNA depletion and stranded method. The ribosomal RNA was depleted from total RNA using the rRNA Removal Kit following manufacturers instruction. RNA was then fragmented into 250~300 bp fragments and reverse-transcribed into cDNA subsequently. Remaining overhangs of double-strand cDNA were converted into blunt ends via exonuclease/polymerase activities. Once the insert size and concentration of the library was identical, the samples can then be subjected for sequencing on Illumina NovaSeq 6000 platform. The metatranscriptome sequencing utilized paired-end (PE) sequencing with 150 base pair (bp) read lengths.