Protocol

Isolation of Individual Cells and Tissues from Electroporated Sea Squirt (Ciona) Embryos by Fluorescence-Activated Cell Sorting (FACS)

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

¹ Present address: Center for Developmental Genetics, Department of Biology, Faculty of Arts and Science, New York University, New York, NY 10003, USA.

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

INTRODUCTION

This protocol describes the dissociation of electroporated sea squirt (Ciona) embryos and the subsequent isolation of individual cells, cell lineages, and tissues, using fluorescence-activated cell sorting (FACS). Sorted samples can be used for RNA extraction and reverse transcriptase polymerase chain reaction (RT-PCR) or microarray analysis. Alternatively, dissociated samples can be used for the subsequent culturing and/or chemical treatment of isolated embryonic cells. This method permits the identification of effector genes responsible for changes in cell shape, polarity, or motility in a given tissue.