2.5. Method 2
The major challenge for the extraction of Cryptosporidium DNA is to lyse the thick wall of oocysts. Since pretreatment steps like snap-freeze and thawing facilitate rupture of the oocyst wall, we have employed the same using Liquid nitrogen (-196C) and incubation at 70C respectively, instead of the sonication step used in the previous method. The remaining steps/procedure performed was similar to method 1.