smRNAseq of A. thaliana Col-0 ecotype in flower buds
Total RNA was extracted from  5-day old seedlings (above ground tissue, grown on soil) or flower bud tissue (young unopened buds) using Trizol (Invitrogen) and treated with RNAse free DNAse (Quiagen)  and total RNA was prepared for ether mRNA and small-RNA (sRNA) sequencing. Library preparation was done using the Novogene in house protocol on the Illumina NovaSeq platform. For mRNA:  was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For sRNA:  and 5 adaptors were ligated to 3 and 5 end of small RNA, respectively. Then the first strand cDNA was synthesized after hybridazition with reverse transcription primer. The double-stranded cDNA library was generated through PCR enrichment. After purification and size selection, libraries with insertions between 18~40 bp were sequenced with SE50Total RNA was extracted from  5-day old seedlings (above ground tissue, grown on soil) or flower bud tissue (young unopened buds) using TRIzol (Invitrogen) and treated with RNase free DNAse (Qiagen)  and total RNA was prepared for either mRNA and small-RNA (sRNA) sequencing. Library preparation was done using the Novogene in-house protocol on the Illumina NovaSeq platform. For mRNA:  total RNA was purified using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis. The library was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For sRNA: 3 and 5 adaptors were ligated to 3 and 5 end of small RNA, respectively. Then the first strand cDNA was synthesized after hybridazition with reverse transcription primer. The double-stranded cDNA library was generated through PCR enrichment. After purification and size selection, libraries with insertions between 18~40 bp were sequenced with SE50