The use of second derivative plots for the determination of mol% guanine plus cytosine of DNA by the thermal denaturation method

Sly L.I., Blackall L.L., Kraat P.C., Tian-Shen T. and Sangkhobol V. (1986) The use of second derivative plots for the determination of mol% guanine plus cytosine of DNA by the thermal denaturation method. Journal of Microbiological Methods, 5 3-4: 139-156. doi:10.1016/0167-7012(86)90009-6


Author Sly L.I.
Blackall L.L.
Kraat P.C.
Tian-Shen T.
Sangkhobol V.
Title The use of second derivative plots for the determination of mol% guanine plus cytosine of DNA by the thermal denaturation method
Journal name Journal of Microbiological Methods   Check publisher's open access policy
ISSN 0167-7012
Publication date 1986-01-01
Sub-type Article (original research)
DOI 10.1016/0167-7012(86)90009-6
Volume 5
Issue 3-4
Start page 139
End page 156
Total pages 18
Subject 1305 Biotechnology
2402 Applied Microbiology and Biotechnology
2404 Microbiology
Abstract Procedures for the estimation of thermal denaturation temperatures for the determination of the mol% G+C base composition of bacterial DNA have been examined with a view to simplifying the procedure and taking advantage of developments in microprocessor controlled spectrophotometers. Thermal denaturation profiles of DNA from 40 strains with base compositions ranging from 35 to 70 mol% G+C were studied. Thermal denaturation temperatures were determined by the conventional Tm method and by using second derivative plots to determine the temperature of inflection (Ti). The results showed that there is a close linear correlation between mol% G+C values calculated using Tm or Ti as the thermal denaturation temperature, provided the thermal denaturation temperature of the internal reference DNA is estimated in the same way. Actual Ti values were approximately 0.3°C higher than corresponding Tm values but the increase was consistent across the range of DNA samples studied. The results also showed that the error introduced by disregarding correction for the effect of thermal expansion was not consistent across the range of DNA samples studied. However, the error was small and in practical terms could be neglected.
Keyword DNA
DNA thermal denaturation
mol% guanine plus cytosine
Tm
Temperature of inflection (Ti)
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import - Archived
 
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