High-throughput genotyping, protein purification featured in Cold Spring Harbor Protocols|
COLD SPRING HARBOR, N.Y. (Mon., Nov. 2, 2009) - The introduction of high-throughput laboratory methods has greatly increased the pace of research into the genetics of complex diseases. Instead of focusing only on one or a few coding variants in a small sample of individuals, the ability to accurately and efficiently genotype many individuals and to cover more of the variation within individual genes has resulted in genetic studies with greater statistical power. Laboratory Methods for High-Throughput Genotyping, from Howard Edenberg and Yunlong Liu at the University of Indiana (http://cmg.iupui.edu/EdenbergLab/ and http://compbio.iupui.edu/people/details/10 ), presents an overview of the commonly used methods for high-throughput single-nucleotide polymorphism (SNP) genotyping for different stages of genetic studies and briefly reviews some of the high-throughput sequencing methods just coming into use. The authors also discuss recent developments in “next-generation” sequencing that will enable other kinds of studies. The article is excerpted from the recently published Genetics of Complex Human Diseases laboratory manual ( http://www.cshlpress.com/link/gchdp.htm ). It is featured in the November issue of Cold Spring Harbor Protocols ( www.cshprotocols.org/TOCs/toc11_09.dtl) and is freely available on the journal's website(http://cshprotocols.cshlp.org/cgi/content/full/2009/11/pdb.top62).
The tandem affinity purification (TAP) procedure was pioneered in yeast for the purpose of purifying and characterizing protein complexes and has since been adapted for use in many organisms, including mammalian systems. The TAP procedure involves two sequential affinity purification steps to avoid non-specific protein interactions, a common problem in identifying proteins in complexes. Bimolecular Affinity Purification (BAP): Tandem Affinity Purification Using Two Protein Baits, from Ezra Burstein and colleagues (http://www.utsouthwestern.edu/utsw/cda/dept24482/files/536923.html ), presents a variation on the TAP procedure in which the affinity moieties are placed on two different proteins of a molecular complex to isolate or detect components present in the complex. This variation, called bimolecular affinity purification (BAP), is suited for the identification of specific molecular complexes marked by the presence of two known components. The article is freely accessible on the website for Cold Spring Harbor Protocols (http://cshprotocols.cshlp.org/cgi/content/full/2009/11/pdb.prot5318).
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Cold Spring Harbor Protocols ( www.cshprotocols.org ) is a monthly peer-reviewed journal of methods used in a wide range of biology laboratories. It is structured to be highly interactive, with each protocol cross-linked to related methods, descriptive information panels, and illustrative material to maximize the total information available to investigators. Each protocol is clearly presented and designed for easy use at the bench—complete with reagents, equipment, and recipe lists. Life science researchers can access the entire collection via institutional site licenses, and can add their suggestions and comments to further refine the techniques.
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