See: Rosengarten RD, et al 2015 for comparative RNAseq on solid support versus pulsing.

All the data are available at dictyexpress.org for your reference

*****
Rafael D. Rosengarten, PhD
Postdoctoral Fellow, Shaulsky lab
Department of Molecular and Human Genetics
Baylor College of Medicine
________________________________________
From: DICTY <[log in to unmask]> on behalf of Kovarik, Michelle <[log in to unmask]>
Sent: Tuesday, May 26, 2015 10:24 AM
To: [log in to unmask]
Subject: [DICTY] development on solid vs in suspension

Hello All,


I am interested in studying the PI3K-PKB pathway in Dictyostelium during the early stages of the social cycle (e.g., first 6 hours). We'll be preparing lysates from the cells for biochemical experiments. We do not plan to do imaging experiments or to look at later stages of development or chemotaxis. I am wondering whether there are advantages to doing the development on a solid support (filter or agar) versus in suspension.


Is development on a filter always preferred for protein recovery? If we do the development in suspension, is it necessary to pulse in cAMP for the cell signaling to be normal? I know that to reach the later stages of development in suspension cAMP must be pulsed in, but I'm not sure whether this makes a difference at earlier times. I've been looking at the development page on the Dictybase website and have the Fey, et al 2007 Nature Protocols paper, but I am wondering if there are any papers comparing cellular signaling during the different development protocols.


I'd appreciate any pointers or references to relevant papers.


Thanks so much,

Michelle

---
Michelle L. Kovarik
Department of Chemistry
Trinity College
300 Summit Street
Hartford, CT 06106


Clement Chemistry Bldg 129
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860-297-5275