The correct option is D Number of nucleotides
Heating causes unwinding of dsDNA into a ssDNA molecule by altering the ordered stacking of the bases and breaking their connecting hydrogen bonds. Single stranded DNA do not obey the Chargaff's rule and thus, A : T and G : C ratio can not be taken into consideration. This ssDNA is then studied by comparing the type and number of nucleotides in single strands. Hence, correct option is D.