In the fourth millennium BCE a cultural phenomenon of monumental burial structures spread along the Atlantic façade. Megalithic burials have been targeted for aDNA analyses, but a gap remains in East Anglia where Neolithic structures were generally earthen or timber. An early Neolithic (3762 – 3648 cal. BCE) burial monument at the site of Trumpington Meadows, Cambridgeshire, U.K. contained the partially articulated remains of at least three individuals. To determine whether this monument fits a pattern present in megalithic burials regarding sex bias, kinship, diet, and relationship to modern populations, teeth and ribs were analysed for DNA and carbon and nitrogen isotopic values respectively. Whole ancient genomes were sequenced from two individuals to a mean genomic coverage of 1.6 and 1.2X and genotypes imputed. Results show that they were brothers from a small population genetically and isotopically similar to previously published British Neolithic individuals, with a level of genome-wide homozygosity consistent with a small island population sourced from continental Europe, but bearing no signs of recent inbreeding. The first Neolithic whole genomes from a monumental burial in East Anglia confirm that this region was connected with the larger pattern of Neolithic megaliths in the British Isles and the Atlantic façade.This work is supported by the Wellcome Trust (Award no. 2000368/Z/15/Z) and St John's College, Cambridge (J.E.R., T.K., R.H., S.A.I, A.R., T.C.O’C., C.C.); the European Union through the European Regional Development Fund (Project No. 2014-2020.4.01.16-0030) (C.L.S.); and the Estonian Research Council personal research grant (PRG243) (C.L.S). E.D’A was supported by Sapienza University of Rome fellowship “borsa di studio per attività di perfezionamento all’estero 2017”