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Frequently Asked Questions: Stable Knockin Cell Line Generation


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Frequently Asked Questions: Stable Knockin Cell Line Generation

  1. What genome editing method do you use in your Cell Line service?
  2. How many guide RNAs do you typically design for a CRISPR Genome Editing Service?
  3. Can you provide off-target analysis report?
  4. How is KI confirmed?
  5. What are the deliverables?

1. What genome editing method do you use in your Cell Line service?

For knockout, point mutation, and DNA insertion (small and large DNA insertion): CRISPR/Cas9.

2. How many guide RNAs do you typically design for a CRISPR Genome Editing Service?

We start with 2 gRNAs and validate them. If no active gRNA is identified, we do another 2 gRNAs. In most projects, two rounds of testing are sufficient.

3. Can you provide off-target analysis report?

Yes, upon customer’s request.

4. How is KI confirmed?

We perform Sanger sequencing of the locus of interest both to determine the nature of the INDEL in the edited genes to guarantee frameshift silencing, and to confirm that the genome is edited. For even greater certainty, we also offer Next Generation Sequencing confirmation, which can detect WT alleles down to 10% of total. Being much more sensitive, NGS screening is ideal for detecting complete editing of multiple copy genes, or polyploid cell lines.

5. What are the deliverables?

- Genetically Engineered Cell Line (up to 2 vials 1x10^6 cells/vial)
- Custom Report

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