Principle / Procedure

The NucleoSpin® gDNA Clean-up procedure is the easiest and fastest way to clean up large-sized DNA from enzymatic reactions, standard “organic” extraction methods or for buffer exchange. Thus, this kit can be used to remove contaminants such as salts, enzymes, nucleotides, labeling reagents, etc. from a DNA sample.
The kit provides a simple and convenient way to purify DNA by selective binding to a silica membrane. The proven silica membrane technology is an effective alternative to the widely used concentrator columns for cleaning and concentrating samples, which were extracted by standard “organic” methods. The procedure follows a simple bind-wash-and-elute procedure (see figure).
In the NucleoSpin® gDNA Clean-up procedure DNA binds to the silica membrane in the presence of a high concentration of chaotropic salt, using Buffer DB. Contaminations are removed in a double wash step with ethanolic buffer. Pure DNA is finally eluted under low ionic strength conditions with slightly alkaline buffer DE (5 mM Tris/HCl, pH 8.5) or water.


NucleoSpin® gDNA Clean-up procedure 

Not recommended for gel or PCR clean-up! For this application please use NucleoSpin® Gel and PCR Clean-up. 

Application data

High recovery of large DNA fragments
DNA fragments of different sizes with and without QIAzol® / TRIzol® contamination (1 %) were purified in triplicate with NucleoSpin® gDNA Clean-up (elution volume 50 µL). DNA fragments of 23 kbp, 9.4 kbp, 6.5 kbp, 4.3 kbp, 2.3 kbp, 2 kbp (Lambda / HindIII Ladder, Fermentas) were analyzed by agarose gel electroporesis.   

Absorption spectrum of the samples before and after purification using NucleoSpin® gDNA Clean-up.

 
High recovery of small DNA fragments

DNA fragments of different sizes with and without QIAzol® / TRIzol® contamination were purified in triplicate with NucleoSpin® gDNA Clean-up (elution volume 50 µL). DNA was analyzed and quantified by Agilent Bioanalyzer 2100.