Protocol

 Copyright 2000-2009 ⓒ NucleoGen Inc. All Rights Reserved.
Contact us
nucleogen@hanmail.net for more information
Tel:031-315-6644     Fax:031-312-2008

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Spin and Vacuum column type

Purified ds or ssDNA in 5 minutes from PCR amplification

Up to 80% recovery of DNA fragments (70 bp-10 kb)

> 99.5% primer removal

Very small elution volumes ; 10 ul

Highly concentrated purified DNA

Fast procedure and easy handling

High reproducible recoveries

No ethanol or isopropanol precipitation

Total procedure is 5 min

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Nucleogen Highcon PCR Purification Kit is designed for fast cleanup of DNA fragments from PCR or other enzymatic reactions. Using silica gel membrane technology, Nucleogen Highcon PCR Purification Kit deliver high-purity DNA suitable for use in all applications and highly concentrated DNA using minimal elution volumes.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

50 preps  … 2315

200 preps  … 2312

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Principle

Nucleogen Highcon PCR Purification Kits contain a silica gel membrane assembly for binding of DNA in high-salt buffer and elution and elution with low-salt buffer or water. The purification procedure removes primers, nucleotides, enzymes, mineral oil, salts, agarose, ethidium bromide, and other impurities from DNA samples. Silica membrane technology eliminates the problems and inconvenience associated with loose resins and slurries. Specialized binding buffers are optimized for specific applications and promote selective adsorption of DNA molecules within particular size ranges.


Procedure

The Nucleogen Highcon PCR Purification System uses a simple bind-wash-elute procedure. Binding buffer is added directly to the PCR sample or other enzymatic reaction, and the mixture is applied to the Nucleogen Highcon PCR Purification spin column. Nucleic acids adsorb to the silica gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in all subsequent applications.


Applications

DNA fragments purified with the Nucleogen Highcon PCR Purification Kits are ready for direct use in most applications, including:

Automated fluorescent sequencing, including capillary sequencing
Radioactive sequencing
Microarray analysis
Ligation and transformation
Restriction digestion
Labeling
Microinjection
PCR
In vitro transcription

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

             a    b    M    1    2

 

 

 

 

 

 

 

- 1100 bp

 

 

 

 

- 94 bp

Analysis of PCR products before (a) and after (b) purification with the Nucleogen PCR Purification Kit(1312).
DNA fragments after extraction (1):Nucleogen PCR Purification (1312), (2):Nucleogen Highcon PCR Purification (2312).
Sample were analyzed on a 1.2% TAE agarose gel.
M:markers

 

 

 

Kit Contents

Highcon PCR Spin Columns, Collection Tubes, PCR Purification Buffer, Wash C solution, Elution Buffer