I have BC1F2, the second generation of backcrossed M2 lines from EMS suppressor screen. All candidate lines are recessive for the parental phenotype. Now, we decided to use NGS for identification of causative mutations according to James at al. 2013. I am going to send for each M2 line the bulk sample of its BC1F2 genomes always from 50 plants. I am not sure if I should send more samples for each of BC1F2 lines?