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cell cloning by limiting dilution

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cell cloning by limiting dilution

Postby lalehr » Fri Sep 04, 2009 10:22 am

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
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Postby jyaron » Sat Sep 05, 2009 10:26 pm

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.
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Re: cell cloning by limiting dilution

Postby jappy » Wed Sep 23, 2009 10:27 pm

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.
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Re: cell cloning by limiting dilution

Postby CellSpecific » Thu Sep 24, 2009 12:48 pm

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.

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http://www.cellspecific.com

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Re: cell cloning by limiting dilution

Postby anubt22 » Mon Sep 06, 2010 5:02 am

jappy wrote: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.
.


Dear Friend,
As I am awair that you have very good hands-on experience of cell cloning by limiting dilution. I have first time planned to clone mouse fibroblast cells by this method. I need you expert guidance. Please send me the protocol for this. And your critical suggessions.

Waiting for your reply.

Thanks

With Best Regards
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