Preliminary dielectric measurement and analysis protocol for determining the melting temperature and binding energy of short sequences of DNA in solution.
Measurement of the real dielectric constant of bulk buffer solutions containing short sequences of DNA as a function of temperature through the DNA melting or denaturiztion transition can be used to determine melting temperatures, T(m), and to estimate the binding energy of the complimentary strands. We describe a preliminary dielectric measurement and analysis protocol to determine these parameters and its application to two known short sequences. The relative real dielectric constant for the bulk solutions was determined over the frequency range of 50 Hz-20 kHz and temperature range of <40-65 degrees C. The measurements were performed on dilute solutions and utilized low electric field strengths. Based on fits to the data by modified sigmoid functions, the melting temperatures, width of transition, and binding energy for the two sequences in solution were estimated. It was observed that the order of the transition appeared to be second order. The results were then compared against predictions of a number of models from the literature that provide theoretical estimates for the melting temperatures of known short sequences of DNA.
Spillman
William B
WB
Department of Biomedical Sciences and Pathobiology, Virginia/Maryland Regional College of Veterinary Medicine, Virginia Tech, Blacksburg, VA, USA. wspillma@yahoo.com
Asmatulu
Ramazan
R
Jullian
Christelle F
CF
Geist
Brian
B
Claus
Richard O
RO
Robertson
John L
JL
D000465:Algorithms; D004247:DNA / Q000737:chemistry*; D004563:Electrochemistry / Q000295:instrumentation / Q000379:methods*; D009691:Nucleic Acid Denaturation; D012996:Solutions / Q000737:chemistry; D013816:Thermodynamics; D044366:Transition Temperature
D012996:Solutions; D004247:DNA