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- W2040390968 abstract "Abstract It is generally accepted that DNA conserves its secondary structure after a freeze-thaw cycle. A negligible amount of degradation occurs after this procedure. Degradation becomes appreciable only after multiple cycles of freezing and thawing. In this study, we have found that a single freeze-thaw cycle in alkaline medium (pH ≥ 10.8) gives rise to denaturation of calf thymus DNA, although the melting temperature of intact DNA in the solution used for the freeze-thaw experiments is higher than 60 °C. The degree of denaturation is almost independent of the regime of freezing. The melting curve obtained after DNA is frozen at −2 °C and then thawed is almost the same as after a freezing carried out in liquid nitrogen (-196 °C). However, incubation in the same solution at 0 °C for 24 hours without freezing does not give rise to any denaturation. The degree of denaturation caused by freezing increases with pH (if pH ≥ 10.8) and decreases with Na2CO3 concentration at fixed pH and [Na+], although Na2CO3 decreases the melting temperature of intact DNA. A preliminary treatment of DNA with cisplatin or transplatin (0.01 Pt atoms per nucleotide) gives rise to a full recovery of the DNA secondary structure after freezing and thawing similar to what occurs after heating DNA to 100 °C and quick cooling. Possible mechanisms that may cause DNA denaturation during a freeze-thaw cycle in alkaline medium are discussed." @default.
- W2040390968 created "2016-06-24" @default.
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- W2040390968 date "2009-02-01" @default.
- W2040390968 modified "2023-10-06" @default.
- W2040390968 title "DNA Denaturation Under Freezing in Alkaline Medium" @default.
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- W2040390968 doi "https://doi.org/10.1080/07391102.2009.10507267" @default.
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