Dilution cloning

The limiting dilution cloning ( to limiting dilution cloning German ) is a method for separating a clone. With the clone, a monoclonal cell culture started.

Principle

The limiting dilution cloning based on the dilution of a suspension of cells in several dilutions up to the limiting dilution, statistically only one or no cells are present in a dilution step in the. By culturing these isolated single cell all the resulting daughter cells are genetically identical clones. In animal cells, are often the simultaneous use of feeder cells is required. Tissue cells as adherent cells have cell-cell contacts, which are released, for example, with trypsin prior to singulation.

Cloning using the limiting dilution is carried out for separation of cells, such as Hybridomata in transgenic organisms after cloning, wherein the induced pluripotent stem cells, in parasites, in T cells, or macrophages.

Alternatives to the isolation by means of the limiting dilution is, for example, FACS, MACS, or a manual isolation under a microscope with microcapillaries.