RNA-Seq of leaf
Treat total RNA with mRNA enrichment or rRNA removal. mRNA enrichment uses magnetic beads with OligodT to capture mRNA. rRNA removal uses DNA probes to hybridize with rRNA, then digests the DNA/RNA hybrids with RNase H, and removes the DNA probes with DNase I. Fragment the RNA and reverse transcribe it with random primers to make double-stranded cDNA. End-repair the cDNA, add a phosphate to the 5' end, and create a 3' overhang with an "A". Then, ligate an adapter with a 3' overhang "T". Amplify the ligated products with PCR using specific primers. Denature the PCR products to single strands and circularize them with a bridging primer to form a single-stranded circular DNA library.
SUB14542186
Leaf1