Please cite as: CSH Protocols; 2006; doi:10.1101/pdb.prot4030

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 Genomic DNA
Right arrow Electrophoresis, general
Right arrow Pulsed Field Gel Electrophoresis
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?
Legend icon

protocolProtocol

Preparation of DNA for Pulsed-field Gel Electrophoresis: Isolation of DNA from Mammalian Cells and Tissues

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

Genomic DNAs from mammalian cells are prepared for pulsed-field gel electrophoresis by lysing cells in situ in an agarose plug. Following digestion with an appropriate restriction enzyme, the plug is loaded directly into the well of a pulsed-field gel or it can be melted before loading.


MATERIALS

Cell or tissue sample

This protocol describes methods for dealing with cultured cell lines, fresh or frozen tissue samples, or white blood cells.

recipe EDTA (0.5 M, pH 8.0)

recipe L buffer

recipe L buffer with proteinase K and Sarkosyl

Amend the L buffer to a final concentration of 1% (w/v) Sarkosyl. Store L buffer with Sarkosyl . . . [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    What's this?