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![You are currently in the section Services > Development > Cloning Navigation - Development - Cloning](https://www.covalab.com/media/wysiwyg/illustrations/website/navigation_2_5.png)
Seeding of selected hybridomas
![Seeding the selected hybridomas using the limiting dilution method Seeding the selected hybridomas](https://www.covalab.com/media/wysiwyg/pictures/monoclonal/cloning_plate_wide.jpg)
At this stage, the selected hybridomas are still polyclonal. To isolate clones from each other, each selected hybridoma is seeded into 96-well plates using the limiting dilution method, and cultured for two weeks.
Identification of growing clones
![Identification of growing clones Screening of growing cell lines](https://www.covalab.com/media/wysiwyg/pictures/monoclonal/screening.jpg)
A visual control is then performed to check for isolated cell clusters which correspond to one single clone. After another week of growth, the supernatant of cell cluster-containing wells are assayed by ELISA to identify the clones which specifically produce the expected antibody. Based on our results, we select up to six positive clones per hybridoma.
Confirmation of antibody production
![ELISA on the supernatant from growing clone-containing wells ELISA](https://www.covalab.com/media/wysiwyg/pictures/corporate/elisa.jpg)
The cells from the corresponding wells are then transferred into a 24-well culture plate and expanded for one week. The supernatants are then assayed again to check for stable antibody production. We also ship to you 1 mL of each supernatant to run tests in your specific conditions.
It's up to you!
![You choose which clone you wish to keep for each hybridoma Validation](https://www.covalab.com/media/wysiwyg/illustrations/miscellaneous/green_tick.jpg)
Based on your results, you select one final clone for each selected hybridoma. The cells are then expanded in 25 cm2 flasks for freezing. A complete isotyping is performed to determine the class and subclass of the antibody as well as the isotype of the light chain.