PUBLICATION

MultiFRAGing: Rapid and Simultaneous Genotyping of Multiple Alleles in a Single Reaction

Authors
Petree, C., Varshney, G.K.
ID
ZDB-PUB-200225-14
Date
2020
Source
Scientific Reports   10: 3172 (Journal)
Registered Authors
Varshney, Gaurav
Keywords
none
MeSH Terms
  • Alleles*
  • Animals
  • Base Sequence
  • Genotyping Techniques*
  • INDEL Mutation/genetics
  • Reproducibility of Results
  • Zebrafish/genetics*
PubMed
32081936 Full text @ Sci. Rep.
Abstract
Powerful and simple, RNA-guided CRISPR/Cas9 technology is a versatile genome editing tool that has revolutionized targeted mutagenesis. CRISPR-based genome editing has enabled large-scale functional genetic studies through the generation of gene knockouts in a variety of model organisms including zebrafish, and can be used to target multiple genes simultaneously. One of the challenges associated with the large scale application of this technique to zebrafish is the lack of a cost-effective method by which to identify mutants. To address this, we optimized the high-throughput, high-resolution fluorescent PCR-based fragment analysis method to develop MultiFRAGing - a robust and cost-effective method to genotype multiple targets in a single reaction. Our approach can identify indels in up to four targets from a single reaction, which represents a four-fold increase in genotyping throughput. This method can be used by any laboratory with access to capillary electrophoresis-based sequencing equipment.
Genes / Markers
Figures
Show all Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping