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

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

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

Economical Kits

No phenol/chloroform extraction

Excellent reproducibility

Total procedure is < 25min

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Nucleogen 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 Gel Extraction Kit deliver high-purity DNA suitable for use in all applications.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

50 preps  ……… 5215

200 preps  ……… 5212

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Principle

Nucleogen 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 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 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 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

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

   

      M         1          2         3

 

 

 

 

 

 

 

 


2.0 kb -
1.2 kb -
0.8 kb -

0.4 kb -


0.2 kb -

0.1 kb -

 

DNA fragments (size indicated) before extraction with Nucleogen Gel Extraction Kit (1212) and after extraction.
Lane M: markers ; lane 1 – 3 : after extraction.
Sample were analyzed on a 1.5% Agarose (agarose-1000, Invitrogen) gel in TBE buffer

 

 

Kit Contents

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