Abstract
The introduction of CRISPR-Cas9 technology for targeted mutagenesis has revolutionized reverse genetics and made genome editing a realistic option in many model organisms. One of the difficulties with this technique is screening for mutations in large numbers of samples. Many screening approaches for identifying CRISPR-Cas9 mutants have been published; however, in practice these methods are time consuming, expensive, or often yield false positives. This report describes a PCR-based screening approach using non-denaturing PAGE. This approach does not depend on the formation of heteroduplexes and reliably detects changes as small as 1 base-pair (bp) in nucleic acid length at the target site. This approach can be used to identify novel mutations and is also useful as a routine genotyping method.
Original language | English (US) |
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Pages (from-to) | 275-278 |
Number of pages | 4 |
Journal | BioTechniques |
Volume | 64 |
Issue number | 6 |
DOIs | |
State | Published - Jun 2018 |
Keywords
- Animals
- CRISPR-Cas Systems/genetics
- DNA/analysis
- Genotyping Techniques/methods
- Mutation/genetics
- Native Polyacrylamide Gel Electrophoresis/methods
- Polymerase Chain Reaction/methods
- Zebrafish/genetics