High Pure PCR Product Purification Kit

Nucleic acids bind to the surface of the glass fiber fleece in the presence of a chaotropic salt (guanidine HCl). This allows the High Pure filter tube to specifically immobilize nucleic acids (both DNA and RNA) while they are freed of contaminants.

Capacity: The High Pure SpinFilter Tubes hold up to 700 μl sample volume.
Volume: 100 μl
Typical samples include:

• Products from amplification or cDNA synthesis reactions
• Enzymatically treated DNA
• DNA from agarose slices
• Dilute nucleic acid solutions
• RNA from transcription reactions

The High Pure PCR Product Purification Kit efficiently and conveniently isolates PCR products from amplification reactions and purifies nucleic acids from other modification reactions. It is also recommended for the purification of cDNA.

The High Pure PCR Product Purification Kit is designed for the preparation of concentrated, purified DNA, and can be used directly for most molecular biology applications:

• Labeling
• Sequencing
• Cloning
• Restriction enzyme digests
• Alkaline phosphatase treatment
• Kinase reactions

The kit eliminates primers, mineral oil, salts, unincorporated nucleotides, and thermostable DNA polymerases, which may inhibit subsequent enzymatic reactions. It can also be applied to concentrate dilute nucleic-acid solutions. Use one kit for a variety of applications.

The fast and simple High Pure protocols use a tabletop centrifuge to bind, wash, and elute the reaction product down to 10 μl (micro format) in as little as 10 minutes. The procedure conveniently eliminates a concentration step, and is ideal for downstream applications such as labeling, sequencing, cloning, ligation, or amplification using PCR.

• Quickly purifies multiple PCR products

in <10 minutes, and purify DNA from agarose gel slices in <20 minutes.

• Efficiently recover DNA fragments

>100 bp in length, and concentrate dilute nucleic acid solutions.

• Minimize DNA loss

with a kit that removes contaminants without precipitation or other handling steps that degrade DNA.

• Eliminate the use of hazardous organic compounds

such as cesium chloride, phenol, chloroform, and ethidium bromide

• Binding Buffer
• Wash Buffer
• Elution Buffer
• High Pure Spin Filter Tubes (containing glass fiber fleece)
• Collection Tubes

More than 70% recovery is obtained when 10 μg DNA Molecular Weight Marker VIII mixed with 16 μg bovine serum albumin are applied to the High Pure Filter Tubes. Gel electrophoresis of the DNA eluate confirms the complete removal of protein and DNA fragments smaller than 100 bp.

The sample is mixed with a chaotropic salt and applied to the glass fiber fleece in a High Pure Spin Filter Tube. Under the buffer conditions used in the procedure, all nucleic acids (NA) in the sample bind to the glass fleece in the High Pure tube, while contaminating substances (salts, proteins, nucleotides, mineral oil and other contaminants) do not. Brief wash-and-spin steps readily remove these contaminants. Once purified, the NA can be easily eluted in a small volume of low salt buffer.