Size Exclusion Spin Columns: Separating The Wheat From The Chaff

Sunday November 11, 2007
Size Exclusion Spin Columns: Separating The Wheat From The Chaff

The need to separate larger molecules from smaller molecules is a recurring theme in life science research. For example, PCR reactions, restriction digests, ligations, and DNA or RNA labeling reactions all generate nucleic acid products that are, in most cases, larger than the reaction components that could interfere with downstream applications. Similarly, protein solutions frequently require buffer exchange or the removal of unincorporated label, such as biotin or 125I. Fortunately, separating the nucleic acid or protein products of interest from smaller contaminants is a simple matter with size exclusion spin columns. With sample volumes ranging from 10 µl to 0.5 ml, finding a spin column to fit your specific application is never a problem. Size exclusion spin columns come ready to use and are preloaded with a variety of buffers that are optimized for different applications. Some spin columns are recommended only for nucleic acids, some only for proteins, and a few are suitable for both nucleic acids and proteins. Size exclusions range from 10 to 200 bp for nucleic acids and from 6 to 12 kDa for proteins. These are the numbers you’ll want to keep in mind when choosing a spin column so you don’t lose your molecule of interest. Just make sure your nucleic acid or protein is larger than the size exclusion, also called the molecular weight cutoff (MWCO), and you’re all set. Since most size exclusion spin columns on the market today have product recoveries of over 90% and can remove over 95% of contaminating small molecules, you really can’t wrong with a spin column from any of the vendors on the Biocompare pages. Below is a short list of size exclusion spin columns to get you started.

Michael Campa, PhD
Assoc. Research Professor of Radiology
Duke University Medical Center

Get Quote For All Ambion GE Healthcare, formerly Amersham Biosciences Roche Applied Science Thermo Scientific Pierce Protein Research Products
Company Ambion GE Healthcare, formerly Amersham Biosciences Roche Applied Science Thermo Scientific Pierce Protein Research Products
Item NucAway™ Spin Columns illustra MicroSpin S-200 HR Columns Quick Spin Columns for biotinylated RNA purification; Sephadex G-50, fine (Available only in the U.S.) Protein Desalting Spin Columns
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Description Ambion's NucAway™ Spin Columns offer a fast and convenient method for the removal of unincorporated nucleotides and salts after probe synthesis reactions. Simply rehydrate the column, centrifuge to remove the interstitial fluid, add the probe to the top of the column, and centrifuge again to elute the probe...
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illustra MicroSpin™ S-200 HR Columns
  • For rapid purification of labeled single-stranded or double-stranded DNA fragments > 50 bases in length in a volume of 51 to 100 µl using spin-column chromatography.

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Quick Spin Columns are ready-to-use disposable column units designed to quickly and efficiently remove unincorporated precursors from RNA labeled with biotin by a polymerization or photobiotin reaction...
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Pierce Protein Desalting Spin Columns are designed to desalt or exchange buffer of protein samples with volumes from 30 to 120 µl. Protein Desalting Spin Columns have exceptional desalting...
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Catalog Number AM10070 27-5120-01 3117464001 89849
Quantity 30 50 20 columns 25
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Applications removal of unincorporated nucleotides and salts after probe synthesis reactions purification of labelled single-stranded or double-stranded DNA fragments > 50 bases in length in a volume of 51-100 µl biotinylated RNA purification desalt or exchange buffer of protein samples with volumes from 30 to 120 µl
Buffer Unbuffered Request Info SSC Tris Buffer
MW Exclusion Request Info Request Info < 72 bp 7 kD
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  Ambion GE Healthcare, formerly Amersham Biosciences Roche Applied Science Thermo Scientific Pierce Protein Research Products

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