Neoseiulus californicus is widely used as an effective biocontrol agent of spider mites. In this study, the complete mitochondrial genome sequence of N. californicus was determined using Oxford Nanopore sequencing technology. The complete mitochondrial genome is 21,318 bp in length and contains 13 protein-coding genes, 2 ribosomal rRNA genes and 22 transfer RNA
genes. Its AT content is 78.4%. All start and stop codons of the protein-coding genes are canonical, except for the missing stop codon for the cox3 gene. The control region was polymorphic in length between the sublines of N. californicus due to variable number of direct repeats. The mitogenome presented in this paper contributes to the study of the genetic structure of N. californicus biocontrol populations.