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soft agar assay


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#1 zhu

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Posted 31 August 2010 - 08:24 AM

Hi, everybdy!
I've done soft agar assay for several times. However, I've got a trouble that cells in the top agar cells adhered to the plate unexpectedly a few days after I seeded the cells .

The following was My protocol.
1. Prepare 0.5% base agar layer and store the plate at 4C.let it sit at room temp for 30 min before using.
2. Prepare 0.6% top agarose solution
3. Harvest and count cells
4. Respend the appropriate number of cells in top agar . Final concentration of top agarose solution was 0.3%.
5. Incubate for 10-30 days and feed cells 2 times a week.

Did anybody have experiences?
What's the reason for the phenomenon that cells in the top agar cells adhered to the plate unexpectedly?

#2 zhu

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Posted 06 September 2010 - 07:35 PM

<_<
Nobody konw?

#3 Adrian K

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Posted 06 September 2010 - 08:34 PM

what sort of cells you are using?
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#4 zhu

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Posted 06 September 2010 - 10:15 PM

Thanks for your reply.
I use murine barrow marrow mesenchymal stem cell.
Do you have any suggestions?

#5 Manish Neupane

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Posted 29 January 2012 - 02:09 PM

Hello Zhu, now I am experiencing similar problem i.e. cells in the top agar cells adhered to the plate. Did you find out the solution? Please let me know




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