Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4007

This Protocol
Right arrow Full Text
Right arrow Update/discuss this protocolDiscussion icon
Right arrow Alert me when this protocol is cited
Right arrow Alert me when comments are published
Right arrow Alert me if a correction is posted
Services
Right arrow Similar protocols in this database
Right arrow Alert me to new releases of protocols
Right arrow Save to Personal Folders
Right arrow Download to citation manager
Right arrow Printer-friendly versionPrinter-friendly version
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Sambrook, J.
Right arrow Articles by Russell, D. W.
Right arrow Search for Related Content
PubMed
Right arrow Articles by Sambrook, J.
Right arrow Articles by Russell, D. W.
Related Collections
Right arrow Molecular Biology, general
Right arrow DNA Purification
Right arrow High-Capacity Vectors
Right arrow High-Capacity Vectors, general
Right arrow BACs
Right arrow Vectors
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?
BSN globe

protocolProtocol

Isolation of BAC DNA from Large-scale Cultures

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

The procedure for isolation of BAC DNA is scaled-up to accommodate 500-ml cultures, which, on average, yield 20-25 µg of purified BAC DNA.


MATERIALS

E. coli strain transformed with BAC isolate

Please see Working with Bacterial Artificial Chromosomes. Prepare a fresh overnight culture; please see Step 1.

recipe Alkaline lysis solution I, ice cold

For large-scale preparations of BAC DNA, sterile Alklaine lysis solution I should be supplemented just before use with DNase-free RNase at a concentration of 100 µg/ml.

recipe Alkaline lysis solution II

recipe Alkaline lysis solution III, ice cold

DNA markers for pulsed-field gel electrophoresis

Ethanol

Optional, please see Step . . . [Full Text of this Article]


METHOD


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?