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