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Spiders (Arthropoda: Arachnida: Araneae) are fascinating and most diverse group of air breathing chelicerate arthropods in the animal kingdom. Three different techniques namely- Spreading technique by Traut, chromosomal fixation by Imai and air drying technique are studied for chromosomal analysis of three species of wolf spiders. Chromosome data are reported for three species belonging to families Lycosidae from India. The diploid chromosomes number (2n) as determined from testicular material was 28 in all three species studied and as follows: Hippasa agelenoides: 2n=28 (26+X1X2), Lycosa bistriata 2n= 28 (26+X1X2), Pardosa birmanica: 2n=28 (26+X1X2). Sex determining mechanism is X1X20 type with chromosomes being acrocentric in morphology. The sex elements are unequal and heteropycnotic during meioitic prophase. &nbsp