This work focuses on using biological data as a unique feature to generate e-Signature. DNA, the blue print of life is of unique nature. The signature created using biological data will be difficult to repudiate in the scenario of a legal dispute. Applications of human DNA are not limited to molecular biology, with the advents of fast growing technologies it is possible to inject DNA into e-Signature for positive identification. The proposed methodology uses Signature DNA as a unique biological feature for the registrant. This work has various phases, the first phase includes creating the Signature DNA using hybridized unique DNA segments of the individual (Registrant) which is the unique identification of the user and difficult to duplicate and repudiate. It generates a Bio-Hash of the Signature DNA. The DNA-Hash generated serves for positive identification of the user which computed with the hash of the e-Document and a random value serve as a Bio-Sign (e-Signature) for the e-Document in the second phase. Bio-Sign converted into QR code with a link to the e-Sign service providers website will ensure usability for verification. In the verification phase the verifier scans the QR code which connects to the e-Service provider's web link. The service provider computes and verifies the document and ensures the e-Signature is valid or not to the verifier. If the signer repudiates the signature, positive identification using DNA helps to achieve Non-Repudiation, the last phase. In the scenario of a legal dispute, the registrant cannot repudiate as the authorities can provide positive identification using the DNA Signature for greater assurance. The proposed technique ensures authentication, integrity and Non-Repudiation with Zero knowledge scenario to the verifier.