Haynes Lab:Notebook/CRISPR Editing/2014/08/07: Difference between revisions
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* CRISPR on chromatin: transfections of g21, g22, g27 into Gal4EED/luc cells | * CRISPR on chromatin: transfections of g21, g22, g27 into Gal4EED/luc cells | ||
* Luc-mCh swap | * Luc-mCh swap: building the donor sequence | ||
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* On 8/05/14, cells were grown in 4 mL P/S-free medium to ~80% confluence in a 6-well plate, 3 with 1 μg/mL dox, 3 without dox | * On 8/05/14, cells were grown in 4 mL P/S-free medium to ~80% confluence in a 6-well plate, 3 with 1 μg/mL dox, 3 without dox | ||
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'''Luc-mCh swap: building the donor sequence''' | |||
* Designed nested PCR primers to gradually build homology arms onto mCherry. The resulting donor should... | |||
** Place mCherry in-frame upstream of the luciferase gene | |||
** Add an tag start codon onto mCherry (we had to use KAH154 as a template, where mCherry has no start) | |||
Revision as of 08:47, 7 August 2014
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08/07/2014
CRISPR on chromatin: transfections
Luc-mCh swap: building the donor sequence
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