Extended Data Fig. 2: Overview of the 5′UTR-MaP library preparation strategy. | Nature Biotechnology

Extended Data Fig. 2: Overview of the 5′UTR-MaP library preparation strategy.

From: Identification of conserved RNA regulatory switches in living cells using RNA secondary structure ensemble mapping and covariation analysis

Extended Data Fig. 2

Cells are subjected to in vivo probing, after which poly(A)+ RNA is isolated and fragmented. Fragments are dephosphorylated to remove any endogenous 5′ phosphate and to resolve 2′-3′-cyclic phosphates generated by chemical fragmentation. Capped RNA fragments are decapped, leaving a 5′ phosphate that can be used to ligate a biotinylated adapter. Ligated fragments are captured via streptavidin-coated beads, after which an adapter is ligated to the 3′ end, and library is prepared as per standard MaP protocol.

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