We found many specific steps and conditions for gram-negative bacteria while working with DNeasy Blood & Tissue Kit.\u00a0These details are described in DNeasy Blood & Tissue Handbook (https:\/\/www.qiagen.com\/us\/resources\/resourcedetail?id=6b09dfb8-6319-464d-996c-79e8c7045a50&lang=en). However we decided to describe this protocol step by step pointing out in some critical steps. This protocol was successfully applied while genomic DNA extraction from Shigella spp. and Salmonella spp. strains for whole-genome sequencing.