We use both chirped-pulse Fourier transform and frequency-modulated absorption spectroscopy to study the rotational spectrum of 2-methoxyethanol (CH3OCH2CH2OH) in several frequency regions ranging from 8.7 to 500 GHz. The resulting rotational parameters permitted a search for this molecule in Atacama Large Millimeter/submillimeter Array (ALMA) observations toward the massive protocluster NGC 6334I, as well as source B of the low-mass protostellar system IRAS 16293−2422. A total of 25 rotational transitions are observed in the ALMA Band 4 data toward NGC 6334I, resulting in the first interstellar detection of 2-methoxyethanol. A column density of
cm−2 is derived at an excitation temperature of
K. However, molecular signal is not observed in the Band 7 data toward IRAS 16293−2422B, and an upper-limit column density of 2.5 × 1015 cm−2 is determined. Various possible formation pathways—including radical recombination and insertion reactions—are discussed. We also investigate physical differences between the two interstellar sources that could result in the observed abundance variations.