Hi-
One way is to grow the cells to stationary phase, let them sit there for 1 day, and then dilute them, the polyphosphate in the conditioned medium causes a G2 arrest. This method has variable results, as being at stationary for too long does odd things to the cells, so I've found that this works 1 in 3 times or so. Tto verify the synchronization, you count the cells in the released population once an hour. If there is a steady increase, the synchronization of course didn't work; if the density is flat for 4-6 hours, and then suddenly jumps by a factor of 1.6-2, it worked..... Another way is nocodazole (microtubule disruption), this locks them in M phase, and then you wash the cells to release them into the cell cycle.
best,
Richard Gomer
Thomas Powell '62 Professor of Biology
Texas A&M University
ILSB MS 3474
301 Old Main Drive
College Station, TX 77843-3474
979 458 5745
________________________________________
From: DICTY [[log in to unmask]] on behalf of Carl Franck [[log in to unmask]]
Sent: Thursday, September 15, 2016 8:33 AM
To: [log in to unmask]
Subject: [DICTY] Please advise as to how to at least partially select Dicty in suspension in some phase of the cell cycle
Dear Dicty Community,
We were hoping someone might have a suggestion for how we can to some degree get a suspension culture readily into cell cycle synchronization. Years ago, an anon referee warned that turning room lights on and off matters. Much obliged for any advice! Sincerely, Kimberly Chen and Carl Franck, Ithaca, New York USA
--
Carl Franck
Associate Professor, Physics
Clark Hall E18, 525
Cornell University, Ithaca, NY, USA
website: https://urldefense.proofpoint.com/v2/url?u=http-3A__franckgroup.lassp.cornell.edu_&d=CwIFAg&c=yHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws&r=G0i-xkKvWepiOT01FF_Nx9XkaEEFt5Dttsc3yIePxBPU44aHfHsMfVSlUygwIJiN&m=kPtLlGKrhd_OxZnv5QCGCwOsLS8BoOG4Ruq1RALTMSQ&s=giQ_iNYCOTKqxDKLz_BlnTVrZIYwM7SDk0FMah76k9Q&e= <https://urldefense.proofpoint.com/v2/url?u=http-3A__franckgroup.lassp.cornell.edu_&d=CwMFaQ&c=yHlS04HhBraes5BQ9ueu5zKhE7rtNXt_d012z2PA6ws&r=G0i-xkKvWepiOT01FF_Nx9XkaEEFt5Dttsc3yIePxBPU44aHfHsMfVSlUygwIJiN&m=5bUHbDBF_ljdOt_t8lYFAShaxVqwWaWvxyPThw5Dv_0&s=-1FZdKTM-o-Ac3ooN67zHD6FX9_xUqCtGWdtSA2nqu0&e=>
lab phones: 607-255-5215<tel:607-255-5215> and 255-3562 (long ring for messages)
home phone: 257-6969 (until 10PM)
cell phone: 607-229-7190<tel:607-229-7190> (sometimes unreachable in my lab, if so, please use above lab phone number)
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