Wolfner became interested in biology as a child.[4] She decided to study at Cornell University because it was well known for genetics.[4] During her undergraduate degree she worked in Gerald Fink's laboratory, studying the control of amino acids in yeast, and graduated in 1974.[4][5] She moved to Stanford University for her graduate degree, where she was a doctoral student in the lab of David Hogness.[6] She was one of the first to use recombinant DNA to isolate the genes in Drosophila.[4] Wolfner pioneered the use of cDNA hybridisation to isolate the genes which respond to ecdysone during metamorphosis.[4]