Haynes Lab:Notebook/CRISPR Editing/2014/08/03: Difference between revisions
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* Gal4EED/luc transfection | * Gal4EED/luc transfection | ||
* qPCR on luciferase - primer test | |||
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* Lipofectamine LTX transfection of cells, 6-well plate format | * Lipofectamine LTX transfection of cells, 6-well plate format | ||
* Gal4EED/luc +dox from T-25 | * Gal4EED/luc +dox from T-25 trypsinized and plated in wells 1-3 (4 mL medium) | ||
* Gal4EED/luc (no dox) from T-25 | * Gal4EED/luc (no dox) from T-25 trypsinized and plated in wells 4-6 (4 mL medium) | ||
** Growth medium added back to original flask to grow up remaining cells as back-up | ** Growth medium added back to original flask to grow up remaining cells as back-up | ||
* Transfection carried out as described in the CSH Syn Bio 2014 manual (2000 ng DNA/ 600 μL transfection mix/ 4 mL cell sample) | * Transfection carried out as described in the CSH Syn Bio 2014 manual (2000 ng DNA/ 600 μL transfection mix/ 4 mL cell sample) | ||
# Gal4EED/luc +dox, plasmid gRNA 20 | |||
# Gal4EED/luc +dox, plasmid gRNA 29 | |||
# Gal4EED/luc +dox, mock | |||
# Gal4EED/luc (no dox), plasmid gRNA 20 | |||
# Gal4EED/luc (no dox), plasmid gRNA 29 | |||
# Gal4EED/luc (no dox), mock | |||
* Note: used complete DMEM (with pen/strep) so cell viability might be suboptimal because of this | |||
* 8/4/14 - carefully replaced original transfection medium with DMEM/10% FBS | |||
Revision as of 10:37, 4 August 2014
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08/03/2014
Gal4EED/luc transfection
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