The majority of cases of ALL demonstrate an abnormal karyotype, either in chromosome number or structural changes. Abnormal chromosome number in childhood acute lymphoblastic leukemia defines distinct biological subgroups with a different response to treatment. The tubes are cultured with three different protocols to save time if one protocol failed. Cultures are then harvested, and cells are fixed and chromosome spreads are prepared. Of 25 patient studied, one patient had psudodiploid karyotype, three patients had tetra-ploid karyotype, four patients had low hypo-diploid karyotype, four patients had high hyper-diploid karyotype, five patients had low hyper-diploid karyotype and eight patients had normal karyotype. Some other factors like, Age, Sex, Consanguinity, Hemoglobin, WBC count, and Type of the leukemia cell also have been evaluated. We found excess number of patients having hypodiploid karyotype but still response to treatment protocols were satisfactory. By comparison between 4 different cultures methods, we find direct method to be more efficient for ploidy analysis.