Abstract: Means and methods for preparing double stranded target DNA molecules for sequencing. In embodiments double stranded backbone DNA molecules comprising 5? and 3° ends are provided that are: ligation compatible with 5? and 3? ends of the target DNA; form a first restriction enzyme recognition site when self-ligated; in a form that enables self-ligation. Methods may comprise providing, if not already present, the target DNA with 5? and 3? ends that are in a form that prevents self-ligation and that are ligation compatible with the backbone DNA 5? and 3? ends. Methods may further comprise ligating the target DNA to the backbone DNA in the presence of a ligase and a first restriction enzyme that cuts the first restriction enzyme recognition site, thereby producing at least one DNA circle comprising a backbone DNA molecule and a target DNA molecule. Linear DNA.

Abstract: Means and methods for preparing double stranded target DNA molecules for sequencing. In embodiments double stranded backbone DNA molecules comprising 5? and 3? ends are provided that are: ligation compatible with 5? and 3? ends of the target DNA; form a first restriction enzyme recognition site when self-ligated; in a form that enables self-ligation. Methods may comprise providing, if not already present, the target DNA with 5? and 3? ends that are in a form that prevents self-ligation and that are ligation compatible with the backbone DNA 5? and 3? ends. Methods may further comprise ligating the target DNA to the backbone DNA in the presence of a ligase and a first restriction enzyme that cuts the first restriction enzyme recognition site, thereby producing at least one DNA circle comprising a backbone DNA molecule and a target DNA molecule.

Abstract: The invention relates to the field of cancer. In particular, it relates to the field of immune system directed approaches for tumor treatment, reduction and control. Some aspects of the invention relate to the identification of tumor specific neoantigens, such as those resulting from frameshift mutations or DNA rearrangements. Such neoantigens are useful for developing tumor treatments, such as vaccines or cellular immunotherapies and other means of stimulating a neoantigen specific immune response against a tumor in individuals. A new class of neoantigens, referred to herein as ‘Hidden Frames’, as well as methods of identifying such neoantigens is provided.

Abstract: Means and methods for preparing double stranded target DNA molecules for sequencing. In embodiments double stranded backbone DNA molecules comprising 5? and 3? ends are provided that are: ligation compatible with 5? and 3? ends of the target DNA; form a first restriction enzyme recognition site when self-ligated; in a form that enables self-ligation. Methods may comprise providing, if not already present, the target DNA with 5? and 3? ends that are in a form that prevents self-ligation and that are ligation compatible with the backbone DNA 5? and 3? ends. Methods may further comprise ligating the target DNA to the backbone DNA in the presence of a ligase and a first restriction enzyme that cuts the first restriction enzyme recognition site, thereby producing at least one DNA circle comprising a backbone DNA molecule and a target DNA molecule.