+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Determination of melting temperature and temperature melting range for DNA with multi-peak differential melting curves

Determination of melting temperature and temperature melting range for DNA with multi-peak differential melting curves

Analytical Biochemistry 479: 28-36

Many factors that change the temperature position and interval of the DNA helix-coil transition often also alter the shape of multi-peak differential melting curves (DMCs). For DNAs with a multi-peak DMC, there is no agreement on the most useful definition for the melting temperature, Tm, and temperature melting width, ΔT, of the entire DNA transition. Changes in Tm and ΔT can reflect unstable variation of the shape of the DMC as well as alterations in DNA thermal stability and heterogeneity. Here, experiments and computer modeling for DNA multi-peak DMCs varying under different factors allowed testing of several methods of defining Tm and ΔT. Indeed, some of the methods give unreasonable "jagged" Tm and ΔT dependences on varying relative concentration of DNA chemical modifications (rb), [Na(+)], and GC content. At the same time, Tm determined as the helix-coil transition average temperature, and ΔT, which is proportional to the average absolute temperature deviation from this temperature, are suitable to characterize multi-peak DMCs. They give smoothly varying theoretical and experimental dependences of Tm and ΔT on rb, [Na(+)], and GC content. For multi-peak DMCs, Tm value determined in this way is the closest to the thermodynamic melting temperature (the helix-coil transition enthalpy/entropy ratio).

Please choose payment method:

(PDF emailed within 0-6 h: $19.90)

Accession: 057591574

Download citation: RISBibTeXText

PMID: 25640587

DOI: 10.1016/j.ab.2015.01.018

Related references

Research on a reference substance for determination of melting temperature after Ph. Helv. VI and Ph. Eur. I. 1. General information on crystals, melting and definition of melting point. Pharmaceutica Acta Helvetiae 48(11): 639-653, 1973

Reference compounds for determination of melting temperature according to Ph. Helv. VI and Ph. eur. I. 4. Determination of purity and melting point with differential scanning calorimetry--results and discussion. Pharmaceutica Acta Helvetiae 49(3-4): 102-107, 1974

The pressure effect on the melting temperature and the melting curves of epsilon iron and MgSiO3 perovskite. Eos, Transactions, American Geophysical Union 90.52, Suppl., 2009

Reference substances for determination of melting temperatures according to Ph. Helv. VI and Ph. Eur. I. 3. Determination of melting point and melting range and results. Pharmaceutica Acta Helvetiae 49(2): 57-65, 1974

A self-consistent estimation method of melting condition based on major elements in volcanic rocks; degree of melting, pressure, H2 O content and melting temperature. Eos, Transactions, American Geophysical Union 89.53, Suppl., 2008

Determination of melting temperature for variant detection using dHPLC: a comparison between an empirical approach and DNA melting prediction software. Genetic Testing 6(3): 169-176, 2002

Estimation of the diversity between DNA calorimetric profiles, differential melting curves and corresponding melting temperatures. Biopolymers 105(11): 832-839, 2016

Melting of cross-linked DNA. III. Calculation of differential melting curves. Journal of Biomolecular Structure and Dynamics 16(1): 59-67, 1998

The effect of melting temperature and a detergent additive on the melting behavior of propyphenazone-containing suppositories. Die Pharmazie 43(1): 51-52, 1988

Discovering rare variants by use of melting temperature shifts seen in melting curve analysis. Clinical Chemistry 51(8): 1331-1332, 2005

Investigation of melting and resolidification of Sm1.8Ba2.4Cu3.4Ox in Ba-Cu-O melt at the temperature below its melting point. Journal of Materials Science Letters 19(14): 1253-1254, 2000

Melting of (Mg, Fe)SiO3-Perovskite to 625 Kilobars: Indication of a High Melting Temperature in the Lower Mantle. Science 262(5133): 553-555, 1993

Dehydration melting of tonalitic compositions; pressure-temperature constraints on melting within the continental crust. Terra Abstracts 8, Suppl, 1996

On the relation between melting temperature and seismic parameter Phi for perovskites; melting-point at the mantle/ core boundary. Acta Geophysica Polonica 50(3): 413-422, 2002

Accurate, Large-Scale Density Functional Melting of Hg: Relativistic Effects Decrease Melting Temperature by 160 K. Journal of Physical Chemistry Letters 8(7): 1407-1412, 2017