title: Microfluidic-aided genotyping of zebrafish in the first 48 h with 100% viability.
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.
authors.affiliation: Department of Mechanical Engineering, University of Utah, Salt Lake City, UT, 84112, USA, raheelsamuel@gmail.com.
authors.forename: Raheel
authors.initials: R
authors.lastname: Samuel
authors.forename: Regan
authors.initials: R
authors.lastname: Stephenson
authors.forename: Paula
authors.initials: P
authors.lastname: Roy
authors.forename: Rob
authors.initials: R
authors.lastname: Pryor
authors.forename: Luming
authors.initials: L
authors.lastname: Zhou
authors.forename: Joshua L
authors.initials: JL
authors.lastname: Bonkowsky
authors.forename: Bruce K
authors.initials: BK
authors.lastname: Gale
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D002823:Chorion; D004625:Embryo, Nonmammalian; D004867:Equipment Design; D060005:Genotyping Techniques / Q000295:instrumentation* / Q000379:methods*; D056656:Lab-On-A-Chip Devices; D044085:Microfluidics / Q000295:instrumentation; D016133:Polymerase Chain Reaction / Q000379:methods; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics*