Deal CCP4 member's
Sorry for asking Off topic question.
Please correct me if i am doing wrong measurements. I am determining the
double stranded concentration using the absorbance spectrophotometer
reading at 260A and extinction coefficients for individual NMPs . But
protein to DNA stoichiometry is coming really off in ITC. I am doubting
weather i am using right method to determine the DNA concentration
accurately or i am doing some wrong calculation. I am calculating the DNA
extinction coefficients by simply multiplying the total number of
individual nucleotide by it extinction coefficients which are as follow
A = 15.4 mM-1 cm-1
C = 7.4 mM-1 cm-1
G = 11.8 mM-1 cm-1
T = 9.6 mM-1 cm-1
The formula which i am using to calculate the concentration is
DNA concentration(M) = ( Abs@260-Abs@330)/Molar extinction
coefficients of DNA
I am multiplying this by dilution factor.
