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cell collection for polysome profiling

sucrose gradient

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#1 Angelika



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Posted 15 October 2013 - 12:56 PM



I need to collect cells for polysome profiling. I have two stable cell lines that I could like to compare (control and stable overexpression of my target gene) that grow at different rates. I have been wondering what is the best method for collecting them for polysomes. I have considered several options:


1) seed the same number of cells and collect both cell lines, let's say on Day 3 when they are about 80-90% confluent, depending on the cell line 

2) seed the same number of cells and wait one more day for the slowly growing cell line to reach the same confluency as the other one, so in other words: seed the same number of cells and collect one cell line at Day 3 and the other at Day 4

3) seed different cell numbers at the start to account for the differences in growth rates and collect on the same day when they reach similar confluency.


I will be grateful for your advice. 



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