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 4°C.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?
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4 replies to this topic
#3
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Legendary Graduate Beggar
Posted 06 September 2010 - 08:34 PM
what sort of cells you are using?
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"what doesn’t kill you, makes you stronger"---Goddess Casandra reminds me to be strong
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..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn’t kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
#4
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member
Posted 06 September 2010 - 10:15 PM
Thanks for your reply.
I use murine barrow marrow mesenchymal stem cell.
Do you have any suggestions?
I use murine barrow marrow mesenchymal stem cell.
Do you have any suggestions?
#5
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member
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
