Song:Experimental Design For Brainbow: Difference between revisions
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*To isolate GFP-positive cells by flow cytometric cell sorting | *To isolate GFP-positive cells by flow cytometric cell sorting | ||
*To establish integrated clonal cell line from single cell | *To establish integrated clonal cell line from single cell | ||
*To validate persistent and efficient expression of GFP after passaging and during differentiation | *To validate persistent and efficient expression of GFP after passaging and during differentiation | ||
<b>Aim 2.</b>To introduce inducible Cre into brainbow-integrated cell lines | <b>Aim 2.</b>To introduce inducible Cre into brainbow-integrated cell lines |
Revision as of 17:31, 6 September 2013
Aim 1.To generate cell lines random-integrated with brainbow cassette
- To construct lentiviral vector carrying Brainbow-2.1 cassette
- To package and transfect lentivirus
- To isolate GFP-positive cells by flow cytometric cell sorting
- To establish integrated clonal cell line from single cell
- To validate persistent and efficient expression of GFP after passaging and during differentiation
Aim 2.To introduce inducible Cre into brainbow-integrated cell lines
Aim 3.To establish live cell imaging and automated lineage tracking system
Aim 4.To manually isolate single cell from differentiation culture
Aim 5.To perform single-cell transcriptomic and epigenomic analysis