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Nucleic acid thermodynamics is the study of how temperature affects the nucleic acid structure of double-stranded DNA (dsDNA). The melting temperature (T m) is defined as the temperature at which half of the DNA strands are in the random coil or single-stranded (ssDNA) state. T m depends on the length of the DNA molecule and its specific ...
Melting curve analysis is an assessment of the dissociation characteristics of double-stranded DNA during heating. As the temperature is raised, the double strand begins to dissociate leading to a rise in the absorbance intensity, hyperchromicity. The temperature at which 50% of DNA is denatured is known as the melting temperature. Measurement ...
The C 0 t value is the product of C 0 (the initial concentration of DNA), t (time in seconds), and a constant that depends on the concentration of cations in the buffer. Repetitive DNA will renature at low C 0 t values, while complex and unique DNA sequences will renature at high C 0 t values. The fast renaturation of the repetitive DNA is ...
An example is they separate when heated at a higher temperature than dissimilar sequences, a process known as "DNA melting". [2] [3] [4] To assess the melting profile of the hybridized DNA, the double-stranded DNA is bound to a column or filter and the mixture is heated in small steps.
Left: unchanged DNA strand. Right: DNA strand intercalated at three locations (black areas). In biochemistry, intercalation is the insertion of molecules between the planar bases of deoxyribonucleic acid (DNA). This process is used as a method for analyzing DNA and it is also the basis of certain kinds of poisoning.
Melting temperature may refer to: Melting point , the temperature at which a substance changes from solid to liquid state Melting temperature, the temperature at which a DNA double helix dissociates into single strands (see Nucleic acid thermodynamics )
Several DNA polymerases have been described with distinct properties that define their specific utilisation in a PCR, in real-time PCR or in an isothermal amplification. Being DNA polymerases, the thermostable DNA polymerases all have a 5'→3' polymerase activity, and either a 5'→3' or a 3'→5' exonuclease activity.
Loop-mediated isothermal amplification (LAMP) primers [1] Loop-mediated isothermal amplification (LAMP) product [1]. In LAMP, the target sequence is amplified at a constant temperature of 60–65 °C (140–149 °F) using either two or three sets of primers and a polymerase like Bst Klenow fragment with high strand displacement activity in addition to a replication activity.