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High Throughput Microinjections of Sea Urchin Zygotes
Authors:Nadezda A. Stepicheva  Jia L. Song
Affiliation:1.Department of Biological Sciences, University of Delaware
Abstract:Microinjection into cells and embryos is a common technique that is used to study a wide range of biological processes. In this method a small amount of treatment solution is loaded into a microinjection needle that is used to physically inject individual immobilized cells or embryos. Despite the need for initial training to perform this procedure for high-throughput delivery, microinjection offers maximum efficiency and reproducible delivery of a wide variety of treatment solutions (including complex mixtures of samples) into cells, eggs or embryos. Applications to microinjections include delivery of DNA constructs, mRNAs, recombinant proteins, gain of function, and loss of function reagents. Fluorescent or colorimetric dye is added to the injected solution to enable instant visualization of efficient delivery as well as a tool for reliable normalization of the amount of the delivered solution. The described method enables microinjection of 100-400 sea urchin zygotes within 10-15 min.
Keywords:Developmental Biology   Issue 83   Sea Urchins   microinjection   sea urchin embryos   treatment delivery   high throughput   mouth pipette   DNA constructs   mRNAs   morpholino antisense oligonucleotides
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