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Functional groups at the 5′-terminus have a strong impact on signal strength. pRNA with a 5′-phosphate (lanes 5–8) yielded enhanced signal strength (∼10-fold) compared to the same RNA with a 5′-hydroxyl end (lanes 1–4). Gel electrophoresis: 10% native PAGE, 1× TBE; RNAs were coupled to nylon membranes by EDC crosslinking.
