I'm going to edite caner cell line T47D. My question is about the proper methodology to select cell colony which are correctly edited by CRISPR-Cas9.
The editing will be a knock-in of SNP mutated sequences for the transcription of mutated protein.
Many thanks
Amr Ghanam
Hi Isabella Lombardo
For me, 48 hrs post transfection, cells were grown in a drug resistance medium (5-days), then collect 80% of cells (mixed population of cells), PCR, sequence, and once we confirm the correct mutation (in the mixed population), limited dilution will be the method of choice for selection of single cell (without any resistance antibiotic anymore)
Hopefully this helps
Amr
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