Characterization and implementation of the MarathonRT template-switching reaction to expand the capabilities of RNA-seq

Schematic outline of cDNA synthesis on mRNA using the template switching activity of MMLV RTs. The outline depicts the steps, including oligo(dT) primer annealing, primer extension, nontemplated “CCC” addition, TSO annealing to the “CCC” overhang, and primer extension on TSO. The last three steps are known as template switching reactions, and nontemplated “CCC” addition and the subsequent TSO annealing step are based on hypotheses that have never been proven directly for MMLV RTs.