Monitoring mitotic cell division in DT40 cells - Events
Monitoring mitotic cell division in DT40 cells - Events
Monitoring mitotic cell division in DT40 cells - Events
Create successful ePaper yourself
Turn your PDF publications into a flip-book with our unique Google optimized e-Paper software.
Generation of <strong>DT40</strong> <strong>cell</strong>s stably<br />
express<strong>in</strong>g H2B:GFP prote<strong>in</strong><br />
• To monitor chromosome segregation, we generated stable clones of wt<br />
and Chk1-/- <strong>DT40</strong> <strong>cell</strong>s express<strong>in</strong>g the chromat<strong>in</strong> marker histone 2B fused<br />
to GFP (H2B:GFP prote<strong>in</strong>)<br />
• Chromosomes <strong>in</strong> those <strong>cell</strong>s can be detected by fluorescence microscopy<br />
(green fluorescence)<br />
General Steps:<br />
1. Obta<strong>in</strong> plasmid cod<strong>in</strong>g for H2B:GFP<br />
2. Electroporate plasmid <strong>in</strong>to <strong>cell</strong>s<br />
3. Select <strong>in</strong>dividual clones express<strong>in</strong>g the plasmid (green clones!)<br />
4. Use clones <strong>in</strong> time-lapse microscopy experiments
Change language