cell cloning by limiting dilution

[lalehr]

hi every body
does any body did limiting dilution for creating single cell clones? is it a straightforward way or not?
any help or protocol will be appreciated
best

[jyaron]

I'm not sure I understand your question.. is this a question for cell synchronization? Cells are technical cloning themselves during division.. if you want single cell clones, try a single cell elution and they'll be synchronized too.

[jappy]

I did cell cloning by dilution technique. This technique consists of dilute your cells so that you have 50 to 100 cells per 60 or 100 mm culture plate. Depending on which cell line you grow, you can obtain easily or difficult a colony from a single cell.
You don't tell if cell line you want to clone is adherent or it grows in suspension.
Show me your e-mail adress and I'll send you a standard protocol.

[CellSpecific]

In addition to above post, often one supplements the medium with factors that promote optimal cell growth. This might be as simple and straight forward as using fetal bovine serum as oppose to normal bovine serum. You might also want to plate/grow under higher serum concentration (e.g., 20%). You'll need to play with the conditions and design one that's appropriate for the cell type you're working with.

Jay Dela Cruz, Ph.D.

CellSpecific
http://www.cellspecific.com

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