Optimizing RNA quality in cryopreserved tissues without preservatives: impact of preservatives, thawing methods and tissue aliquot sizes

Preservatives, tissue aliquot sizes, and thawing methods significantly impact the RNA quality of frozen tissues originally stored without preservatives. Key recommendations include (1) adding RNALater during thawing, (2) thawing on ice for small aliquots (≤ 100 mg) or at -20 °C for larger samples, and (3) minimizing freeze‒thaw cycles, despite observed variations among species.