In brief, we deliver RNPs containing SpCas9, donor template and the modified sgRNA into the cells using Nucleofection. Post cell recovery, we extract the genomic DNA, PCR amplify the edited site, sequence the site using Sanger sequencing and analyze the data using ICE software. For clonal projects, we single-cell isolate and expand clones in 96-well plates and re-analyze the clones once they have reached an appropriate size.
Articles in this section
- What are Advanced Cell Projects?
- What kind of genetic modifications can you perform?
- What types of cells can you edit?
- What do I receive as part of an Advanced Cell Project?
- Can you deliver heterozygous clones?
- What are the levels of homologous recombination/knock-in efficiencies that you can achieve?
- Do I need to design the guide RNAs or donor templates?
- What is your turnaround time?
- I have already attempted to generate this modification and failed. Why would you succeed?
- What if I do not know if my cell line is capable of single cell expansion?