The establishment of in vitro culture from shoot tip explants (meristemetic tissue) of grapes was investigated through tissue culture technique. These explants were collected from gene bank of Institute of Agricultural Biotechnology and Genetic Resources (IABGR), National Agricultural Research Centre (NARC), Islamabad, Pakistan. Fifteen accessions of grapes were surface sterilized and tested on 75% MS media for germination and initial growth parameters. Accession No. 020017 (Dakh-1) exhibited highest viability (100%), shoot length (4.12 cm) -1 and nodes plant (3.8). Moreover, it was found that response of cultures to different treatments was dependent both on accession and treatment duration. In conclusion, this protocol proved to be useful in optimizing the dose and duration of the treatment of grape explants with the surface disinfectant.