Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3289

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Screening an Unamplified Cosmid Library by Hybridization: Plating the Library onto Filters

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 15% of the full text of this article appears below.


INTRODUCTION

Unamplified cosmid libraries are plated at high density onto 150-mm nitrocellulose or nylon filters and screened by hybridization.


MATERIALS

E. coli plating bacteria

Use the appropriate strain (e.g., XL1-Blue, ED8767, NM554, DH5{alpha}MCR).

Bacteriophage {lambda} packaging reaction

Please see Construction of Genomic DNA Libraries in Cosmid Vectors, Step 19.

recipe TB

recipe TB agar plates (150 mm) containing 25 µg/ml kanamycin


METHOD

1. Calculate the volume of the packaging reaction (Construction of Genomic DNA Libraries in Cosmid Vectors, Steps 20-21) that will generate 30,000-50,000 . . . [Full Text of this Article]


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