Isolation of Sea Squirt (Ciona) Gametes, Fertilization, Dechorionation, and Development

Lionel Christiaen; Eileen Wagner; Weiyang Shi; Michael Levine

doi:10.1101/pdb.prot5344

Protocol

Isolation of Sea Squirt (Ciona) Gametes, Fertilization, Dechorionation, and Development

Molecular and Cell Biology Department, University of California, Berkeley, California 94720, USA

↵²Corresponding authors (mlevine{at}berkeley.edu); (lc121{at}nyu.edu).

INTRODUCTION

This protocol is the starting point for most manipulations that are used to study the sea squirt (Ciona) embryo, including in situ hybridization, the microinjection of morpholino oligos, and the electroporation of transgenic DNAs. Ciona eggs and embryos are exquisitely sensitive to even trace amounts of detergent; therefore, it is strongly advised to designate a soap-free workspace for embryo culture. Any solutions that come into contact with embryos should be prepared in absolutely clean glassware with the highest quality water.