Cite as: Cold Spring Harb. Protoc.; 2006; doi:10.1101/pdb.prot4050

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 RNA, general
Right arrow mRNA
Right arrow Electrophoresis, general
Right arrow Electrophoresis of Nucleic Acids, general
Right arrow Electrophoresis of RNA
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

Separation of RNA According to Size: Electrophoresis of RNA through Agarose Gels Containing Formaldehyde

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 15% of the full text of this article appears below.


INTRODUCTION

Separation of RNAs according to size is the first stage in northern blotting and hybridization. The method described in this protocol uses formaldehyde to denature the RNA, ethidium bromide to stain it, and electrophoresis through agarose gels containing 2.2 M formamide to separate the resulting formaldehyde-RNA-ethidium adducts. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O.


MATERIALS

recipe 10x Formaldehyde gel-loading buffer

recipe 10x MOPS electrophoresis buffer

recipe caution Ethidium bromide (200 µg/ml)

recipe caution Formaldehyde

recipe caution Formamide

Purchase or prepare a distilled-deionized preparation of this reagent and store in small aliquots under nitrogen at -20°C.

RNA samples

. . . [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?