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The normal chromosomal pattern in Nili-Ravi buffalo was studied from the lymphocytic culture of 5 each of the female and male animals kept at Livestock Experiment Station, University of Agriculture, Faisalabad. Micro method for culturing lymphocytes was used for chromosome analysis with slight modification. The chromosomal spread trapped at metaphase stage revealed that 2n diploid number of chromosomes in female and male was 50. The female had 48 autosomes and 2 X-chromosomes. The male had also 48 autosomes along with one X and y chromosomes. The number of meta/submetacentric and acrocentric autosomal chromosomes was 10 and 38, respectively in both the male and female animals. The sex chromosomes in male and female animals were acrocentric.