Chez Charles/ biblio/ T4 RNA ligase: substrate chain length requirements. Edit RecentChanges History Preferences Branchable FEBS Lett. 1974 Sep 15;46(1):271-5. Kaufmann G, Klein T, Littauer UZ. T4 RNA ligase: substrate chain length requirements. For circularisation, (pA)8 is the shortest substrate, and the reaction is most efficient with (pA)10. The ligation efficicency then decreases with substrate length.