CloneMedia Semi-Solid Media For rapid cloning of transfected mammalian cell lines Molecular Devices unique range of ready-to-use semisolid CloneMedia products offer the means to clone mammalian cell lines in a rapid, one step protocol (see the workflow right), equally suitable for manual as well as automated use in cell line and antibody development. Semi-solid media facilitate the growth of single cells into discrete, clonal colonies and the subsequent manual or automated isolation of these clones. This process is simpler and faster than alternative methods (such as limiting dilutions) and eliminates the need for subsequent re-cloning in most cases. Using this technique, all potentially valuable clones (transfectants or hybridomas) arising from a transfection or fusion experiment can be plated out in parallel and colonies picked for expansion within days of plating. Prepare Cells Cloning – perform fusion/transfection (Optional; bulk selection in liquid media). Subcloning – establish line in liquid media. Plate Cells Prepare CloneMedia. (Optional; add fluorescent detection agent e.g. CloneDetect). Add cells and dispense into tissue culture plates. Colony Growth Colony growth can be monitored by high speed CloneSelect Imager for objective, quantitative assessment of cell growth and monoclonality verification. The following CloneMedia products are available: CloneMedia-CHO for the cloning of CHO-S cell lines. Available with or without Glutamine. Compatible with DHFRselection of CHO-DG44 cells. CloneMedia-Hybridoma/Myeloma for the cloning of hybridoma or myeloma cell lines. CloneMedia-HEK for the cloning of suspension-adapted, serum-free HEK293 cell lines. If using CloneDetect colonies can be imaged and analyzed on ClonePix System and highest secretors identified. CloneMedia-CHOK1 for the semi-solid cloning of suspension-adapted, serum-free CHOK1 cell lines, also compatible with CHOK1SV. • All CloneMedia products contain appropriate supplements to promote colony growth in semi-solid media for each cell type • They do not contain antibiotics or other selective agents • Supplied as 90 mL in 100 mL bottles for optional addition of up to 10 mL additional components • Products for CHO, CHOK1 and HEK293 are all suitable for chemically-defined growth conditions • CloneMedia Products were developed by Molecular Devices for optimised use with ClonePix™ mammalian colony picking system as well as CloneSelect™ Imager. Pick and expand highest expressing cell lines. Figure 1: Schematic of a typical workow of the semi solid cloning process.

Figure 2: Images of colonies of commonly used cell types raised in semi-solid culture in CloneMedia. Images were captured using Molecular Devices CloneSelect Imager at a suitable time point post plating.A) CHO colonies, serum-free suspension-adapted, in CloneMedia-CHO imaged on day 8 post-plating. B) Hybridoma colonies in CloneMedia-Hybridoma/Myeloma imaged on day 8 post-plating. C) HEK293 colonies, serum-free, suspension-adapted, in CloneMedia-HEK imaged on day 13 post-plating. D) CHO-K1 colonies, serum-free, suspensionadapted, in CloneMedia-CHOK1 imaged on day 9 post-plating Synergy between...