Cite as: Cold Spring Harb. Protoc.; 2007; doi:10.1101/pdb.prot4752

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Dissection of Tightly Adhering Xenopus laevis Tissues by Trypsin Treatment

Hazel L. Sive, Robert M. Grainger, and Richard M. Harland

This protocol was adapted from "Microdissection," Chapter 10, in Early Development of Xenopus laevis, by Hazel L. Sive, Robert M. Grainger, and Richard M. Harland. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2000.


INTRODUCTION

In older Xenopus laevis embryos (late gastrula and beyond), tissues begin to stick to one another and cannot be peeled apart. For assays requiring isolated tissues, it is necessary to separate such tissues enzymatically, which can be readily accomplished by mild trypsin treatment. Embryos are treated singly, or in small numbers, to avoid possible toxic effects of excessive trypsin digestion. This protocol describes a method for the separation of neural tissue from a neural-plate-stage embryo, although the technique can be adapted to many different tissue types. Enzymatically dissected tissue can be used in quantitative gene expression assays or in specification or induction assays.


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