Abstract: 【Objective】 Ostrinia furnacalis is the main pest that causes maize yield reduction in Xinjiang. Ostrinia nubilalis (Hubern) and Ostrinia furnacalis (Guenee) are sister species that are difficult to be identified by morphology. The Yili River Valley in Xinjiang is a mixed occurrence area of the Eurasian corn borer, so it is very important to carry out accurate isolation and identification of the Eurasian corn borer in this area. a non-destructive extraction method for the corn borer will be used to efficiently establish a pure line population, which is very important for the later biological ecology research. 【Methods】 The purpose of this paper is to establish a DNA extraction method without morphological damage, economical, simple, rapid and can meet the subsequent PCR amplification detection technology. Based on the common DNA extraction methods such as screening kit method (KM), protease method (PM and CTAB method (CM), the DNA extraction, target COI sequence amplification and agarose gel detection of corn borer molt were carried out. 【Results】 The screening results showed that the effective extraction method for the molt was treatment 3: protease method. The DNA concentration of the molt extracted by the protease method was the highest, which was (23.03 ± 0.01) ng/μL, but its purity was not high, which was not conducive to subsequent PCR amplification. The average concentration of DNA extracted by treatment 3 was (8.46 ± 0.22) ng/μL, meeting the extraction requirements, and the purity was 1.86, meeting the subsequent PCR amplification experiment. 【Conclusion】 The treatment 3 and protease method mentioned in this paper can be used to extract gDNA from corn borer molt in large quantities, and at the same time to carry out population identification and subsequent experimental research. Both of them have the advantages of no damage to corn borer, low cost, simple steps, avoiding the harm of chloroform and isopropanol to human body, and it has higher amplification efficiency than the protease method.