Lipopolysaccharide (LPS) is a major component of the outer membrane of Gram-negative bacteria. Some lipopolysaccharides are pathogenic endotoxin, which is also important for serotyping. A common method is the complex hot phenol-water extraction, which takes a long time. The LPS extraction kit is based on a robust water/phenol/chloroform extraction technique and is flexible enough to run small (<5 mL) or large cultures (up to 100 mL) with similar yields. This LPS extraction kit provides a new method for extracting LPS with a higher yield, fewer steps, and less time.