Protocol

Isolation of BAC DNA from Small-scale Cultures

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

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INTRODUCTION

BAC DNAs are prepared from 5-ml cultures of BAC-transformed cells by a modification of the standard alkaline lysis method (Preparation of Plasmid DNA by Alkaline Lysis with SDS: Minipreparation). The yield typically varies between 0.1 and 0.4 µg of BAC DNA.

MATERIALS

E. coli strain transformed with BAC isolate

Please see Working with Bacterial Artificial Chromosomes.

Alkaline lysis solution I, ice cold

For large-scale preparations of BAC DNA, . . . [Full Text of this Article]