Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4069

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Construction and Screening of Eukaryotic Expression Libraries

Stage 2: Screening cDNA Libraries Constructed in Eukaryotic Expression Vectors

Joseph Sambrook and David W. Russell

This protocol was adapted from Molecular Cloning, 3rd edition, by Joseph Sambrook and David W. Russell. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2001

The first 100 words of the full text of this article appear below.


INTRODUCTION

Two methods of screening pools of cloned cDNAs are discussed: transfection into mammalian cells and injection into Xenopus oocytes.


MATERIALS

Host cells

Xenopus oocytes are taken from female South African clawed frogs, which are available from several biological supply houses (such as Carolina Biological Supply or Kons Scientific). Difficulties in expressing mRNAs can sometimes be overcome by switching suppliers.

Simian COS cells or derivatives of the 293 line of human embryonic kidney cells are the mammalian hosts of choice for transient expression of cloned cDNAs because of their superior efficiencies of transfection. However, many other cell lines have been used successfully . . . [Full Text of this Article]


METHOD


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