Red clover IFS1 KO expression
mRNA was enriched using oligo(dT) beads, rRNA removed using the Ribo-Zero kit leaving mRNA. The mRNA was fragmented randomly by adding fragmentation buffer, then the cDNA was synthesized by using mRNA template and random hexamers primer, followed by second-strand synthesis buffer (Illumina) , dNTPs, RNase H and DNA polymerase I were added to initiate the second-strand synthesis. Following a series of terminal repair, ligation and sequencing adaptor ligation, the double-stranded cDNA library completed through size selection and PCR enrichment.