Figure 1

Small RNA cloning strategy. (A) SmRNAs are isolated by size from a sample of total RNA. (B) Then, 5'- and 3'-cloning primers (5-CP and 3-CP) are ligated to smRNAs using T4 RNA ligase[15]. Reverse transcriptase converts RNA to cDNA, which can be subsequently (C) concatenated. (D) These long strands of DNA are then ligated into a plasmid[29], transformed into bacteria and plasmids from single colonies isolated for DNA sequencing. (E) The resulting DNA sequence contains the original smRNA sequence flanked by two cloning primers (5-CP and 3-CP).