Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot3990

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Purification of Bacteriophage {lambda} Arms: Centrifugation through Sucrose Density Gradients

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

This method, derived from Maniatis et al. (1978), is used to prepare the arms of any bacteriophage {lambda} vector.


MATERIALS

cautionn-Butanol

Bacteriophage {lambda} DNA

Prepared as described in Extraction of Bacteriophage {lambda} DNA from Large-scale Cultures Using Proteinase K and SDSor Extraction of Bacteriophage {lambda} DNA from Large-scale Cultures Using Formamide.

recipe EDTA (0.5 M, pH 8.0)

Ethanol

Optional, please see Step 5.

recipe Gel-loading buffer IV

recipe caution MgCl2 (1 M)

recipe NaCl (1 M)

recipe caution Sodium acetate (3 M, pH 5.2)

recipe TE (pH 7.6 and pH 8.0)


METHOD

1. This step serves as an optional preliminary sequence of steps that may be performed before Step 3 for . . . [Full Text of this Article]


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