Root lesion nematode (Pratylenchus spp.) is an economically important plant pathogen after root knot and cyst nematodes. The study aimed to isolate and characterize Pratylenchus nematodes from a farm in New Delhi using both morphological and molecular analysis to allow for accurate identification. Soil samples were collected from different crops and nematodes were extracted, isolated, and studied using microscopy as well as analysis of the internal transcribed spacer region of ribosomal DNA.