Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4040

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 Genomic DNA
Right arrow Blots
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

Southern Blotting: Capillary Transfer of DNA to Membranes

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 protocol describes the first stages of Southern blotting: digestion of genomic DNA with one or more restriction enzymes, separation of the resulting fragments by electrophoresis through an agarose gel, and transfer of the denatured fragments to a membrane by downward capillary transfer.


MATERIALS

recipe 6X Gel-loading buffer IV

recipe 6X SSC

6x SSC should be warmed to various temperatures. Please see Steps 11, 12, and 13.

recipe Alkaline transfer buffer

recipe DNA-staining solution

recipe Denaturation solution

Depending on the number of filters to be processed, 1 liter or more may be required.

Genomic DNA

Prepare high-molecular-weight DNA in TE from appropriate cells as described in . . . [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?