Micro RNA sequencing in Bipolaris sorokiniana: Resistant control 2
Small RNA sequencing (smRNA) libraries were constructed using QIAseq miRNA Library Kit (Cat: 331502) protocol following manufacturers instructions. Briefly, 63ng of total RNA was used as starting material. 3 adapters were ligated to the specific 3OH group of micro RNAs followed by ligation of 5 adapter. Adapter ligated fragment was reverse transcribed with Unique Molecular Index (UMI) assignment by priming with reverse transcription primers. cDNA thus formed was enriched and barcoded by PCR amplification (17cycles). The Illumina-compatible sequencing libraries were quantified by Qubit fluorometer. The fragment size distribution of the libraries was analyzed on Agilent 2200 TapeStation. Finally, these libraries were sequenced on Illumina NovaSeq 6000 using SE50 read length and sequencing chemistry.