Fig. 5: Analysis and purification of circRNA using high-performance liquid chromatography (HPLC).
From: Chemical circularization of in vitro transcribed RNA for exploring circular mRNA design
A Samples of linear (p)RNA04 and crude post-circularization mixture containing linear (p)RNA04 and circ(p)RNA04, (RNA04 760 nt) were analyzed using different HPLC methods: ion-exchange (IEC), size-exclusion (SEC), and reversed-phase (RP). B IP-RP-HPLC separation of linear and circRNA05 (578 nt) followed by PAGE analysis of collected fractions. Column and buffers as in A—IP-RP (Supplementary Information). Fractions F1 contains mostly precursor RNA; F2 contains the circular product of high purity; F3 includes linear dimers and concatemers. C Resolution of linear (lower retention time) and circular (higher retention time) RNA06 (598 nt) using IP-RP-HPLC with different ion-pairing reagents: triethylammonium acetate (TEAA), diisopropylammonium acetate (DIPAA), or n-hexylammonium acetate (HAA). A–C For purification/separation conditions please see Supplementary Information.
