FAQ: Does RNA secondary structure impact RNase 4 (NEB #M1284) activity?

For optimal nucleotide sequence mapping coverage, it is recommended to perform RNase 4 digests under denaturing conditions to limit the possible influence of RNA secondary structure. A digest reaction containing a final urea concentration at 1 M, incubated at 37 °C for 1 h, will ensure most RNA is available for endonucleolytic cleavage.

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