Identification of QTLs for resistance to Plasmodiophora brassicae in Brassica oleracea through BSA-seq
Using the Ribo-Zero kit, rRNA was removed from the total RNA samples. Then, fragmentation buffer was added to break the RNA into small fragments. With the fragmented RNA as a template, 6bp random hexamers were used to synthesize the first strand of cDNA. Subsequently, buffer, dNTPs (with dTTP replaced by dUTP), DNA polymerase I, and RNase H were added to synthesize the second strand of cDNA. The synthesized double-stranded cDNA was purified, end-repaired, adenylated, and ligated with sequencing adapters. USER enzyme was then used to degrade the second strand of cDNA containing uracil (U). PCR enrichment was performed, and the PCR products were purified using AMPure XP beads to obtain the final strand-specific library.
SUB14788271
FRRL19H001605-1a