Amplified product of ESR1 gene (rs 9340799) of clinically diagnosed breast cancer patients of Pashtun ethnic group. A, Lanes 1,2,3,4 show amplified product of 524bp. Lane M is 100bp ladder as a marker. B, Digestion of amplified PCR product (524bp) with Xbal restriction enzyme. Lanes 1, 3 and 6 show the samples completely digested Xbal enzyme, producing 2 bands of 297 and 227bp (pretty close to each other that appears as single band). This confirms the presence of G>A polymorphisms and are homozygous for minor allele “A”. Lanes 2, 5 and 7 show the samples incompletely digested by the Xbal restriction enzyme and are thus heterozygous for minor allele “A”, giving 3 bands of 524, 297 and 227bp. Lanes 4 show sample not digested by Xbal enzyme and thus showing a single band of 524bp. This sample is homozygous for major allele “G”. M is a 100bp ladder. Lane 8 is blank. C, amplified product of ESR1 gene (rs2234693) of clinically diagnosed breast cancer patients of Pashtun ethnic group. D, Digestion of amplified PCR product using PvuII restriction enzyme. Lanes 1 to 8, samples are not digested by PvuII restriction enzyme, showing a single band of 451b. Thus all the patients lack rs2234693 (C>T) polymorphism. M is a 100bp ladder used as a marker.