The DNA extraction method must be simple, quick and efficient. Safety, cost and DNA quality must also be considered. Bacillus thuringiensis produces endospores that resist the lysis by usual methods used for other bacteria. SDS - NaOH is mostly used in lysis buffer along with incubations for lengthy time periods. Here we devise a rapid method for DNA extraction from Bacillus thuringiensis using triton X-100 in lysis buffer. After incubation at 60°C for 30 min, the lysate is phenol-chloroform extracted. DNA was precipitated with absolute ethanol, washed with 70% ethanol and dissolved in TE buffer. The entire method did not take more than 1 hour. The DNA extracted by this method was utilized in enzymatic reactions including PCR, restriction analysis and produced good results. This method can also be used for DNA extraction from other species including Gram negative as well as Gram positive isolates.