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• W1488387004 abstract "Publisher Summary This chapter presents a systematic method for extracting the thermodynamic contributions of RNA tertiary structure to the folding and overall stability of any RNA for which the secondary structure is known. The procedure can be applied to RNAs of modest length, up to 100 bases, and yields an approximation of the temperature-dependent RNA-unfolding pathway. Further studies can build on this information to try to identify the specific nucleotides contributing to the tertiary interactions, and to study the thermodynamics of specific ligand interactions. The thermodynamic analysis is based on melting experiments carried out on series of RNAs with different termini and with mutations in key structures. RNA synthesis on a milligram scale by in vitro transcription is a well-established method. The polymerase chain reaction to link a T7 promoter to various segments of a DNA sequence for transcription: this allows a great deal of flexibility in designing a series of RNAs with different 5' and 3' termini. There are two useful methods for following the thermal denaturation of an RNA; they rely on changes in either ultraviolet (UV) absorption or heat capacity on unfolding of the RNA. A problem with the interpretation of UV melting curves of larger RNAs is that it may be necessary to fit more variables than the data can bear, because the hyperchromicity of each transition is an unknown in addition to the thermodynamic parameters." @default.
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• W1488387004 date "1995-01-01" @default.
• W1488387004 modified "2023-09-24" @default.
• W1488387004 title "[13] Melting studies of RNA unfolding and RNA—Ligand interactions" @default.
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• W1488387004 doi "https://doi.org/10.1016/0076-6879(95)59049-8" @default.
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