The mango (Mangifera indica L.) is a common tropical and subtropical fruit and the second main fruit crop in Egypt. In this work, start codon targeted (SCoT) and inter simple sequence repeat (ISSR) markers were employed to study the genetic relationship among twenty-three mango genotypes grown under the El-Giza district conditions. These genotypes consist of four newly introduced cultivars, six new progenies and thirteen closely related mango cultivars which were included as cultivar references for the best definition. Based on the overall SCoT and ISSR data, a total of 52 bands were generated (18 for SCoT and 34 for ISSR) with a total polymorphic percentage equal to 53.84% (44.44% for SCoT and 58.82% for ISSR). Based on the whole set of SCoT and ISSR marker data, the genetic similarity coefficients ranged from 80 to 97% with a mean equal to 88.5% and the first two principal axes explained about 31% of the total genetic variation among the mango accessions. The SCoT and ISSR data revealed that the four newly introduced mango genotypes, namely ‘Aya’, ‘Kasturi’, ‘Maya’ and ‘Omer’, are novel genetic resources based on their genetic profiles, which can be used for future breeding programmes in Egypt.