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

Very small elution volumes ; 10 ul

Highly concentrated purified DNA

Gel extraction from standard or low-melt agarose gels in TAE or TBE buffer

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

Fast procedure and easy handling

High repoducible recoveries

No phenol/chloroform extraction

Total procedure is < 25min

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Nucleogen Highcon Gel Extraction Kit is designed for fast cleanup of DNA fragments from standard or low-melt agarose gels in TAE or TBE buffer. Using silica gel membrane technology, Nucleogen Highcon Gel Extraction Kit deliver high-purity DNA suitable for use in all applications and highly concentrated DNA using minimal elution volumes.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 50 preps  … 2215

 200 preps  … 2212

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Principle

Nucleogen Highcon Gel Extraction Kits contain a silica gel membrane assembly for binding of DNA in high-salt buffer 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 Gel Extraction system uses a simple bindwash- elute procedure. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the Nucleogen Highcon Gel Extraction 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.


pH indicator dye

pH indicator dye in the solubilization and binding buffer allows easy visual determination of optimal pH for DNA adsorption (pH ≤7.5). An incorrect binding-mixture pH can occur if the agarose gel electrophoresis buffer was frequently used or incorrectly prepared. In this case the pH can be easily adjusted by addition of 10 ul 3 M sodium acetate, pH 5.0.


Applications

DNA fragments purified with the Nucleogen Highcon Gel Extraction 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

 

 

DNA fragments before (a) and after (b) extraction with the Nucleogen Gel Extraction Kit(1212). DNA fragments after extraction (2):Nucleogen Highcon Gel Extraction (2212), (1):Nucleogen Gel Extraction (1212).
Sample were analyzed on a 1.2% TAE agarose gel.
M:markers

 

 

Kit Contents

Highcon Gel Spin Columns, Collection Tubes, Gel Extraction Buffer, Wash C solution, Elution Buffer