PUBLICATION

Microfluidic-aided genotyping of zebrafish in the first 48 h with 100 % viability

Authors
Samuel, R., Stephenson, R., Roy, P., Pryor, R., Zhou, L., Bonkowsky, J.L., Gale, B.K.
ID
ZDB-PUB-150317-3
Date
2015
Source
Biomedical Microdevices   17(2): 43 (Journal)
Registered Authors
Bonkowsky, Joshua
Keywords
Zebrafish, Genotyping, Microfluidics
MeSH Terms
  • Animals
  • Animals, Genetically Modified
  • Chorion
  • Embryo, Nonmammalian
  • Equipment Design
  • Genotyping Techniques/instrumentation*
  • Genotyping Techniques/methods*
  • Lab-On-A-Chip Devices
  • Microfluidics/instrumentation
  • Polymerase Chain Reaction/methods
  • Zebrafish/embryology*
  • Zebrafish/genetics*
PubMed
25773537 Full text @ Biomed. Microdevices.
Abstract
This paper introduces an innovative method for genotyping 1-2 days old zebrafish embryos, without sacrificing the life/health of the embryos. The method utilizes microfluidic technology to extract and collect a small amount of genetic material from the chorionic fluid or fin tissue of the embryo. Then, using conventional DNA extraction, PCR amplification, and high resolution melt analysis with fluorescent DNA detection techniques, the embryo is genotyped. The chorionic fluid approach was successful 78 % of the time while the fin clipping method was successful 100 % of the time. Chorionic fluid was shown to only contain DNA from the embryo and not from the mother. These results suggest a novel method to genotype zebrafish embryos that can facilitate high-throughput screening, while maintaining 100 % viability of the embryo.
Genes / Markers
Figures
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping