DESCRIPTION
The GeneJET™ Gel Extraction Kit is designed for rapid and efficient purification of DNA fragments from standard or low-melting point agarose gels run in either TAE or TBE buffer.
The kit utilizes a proprietary silica-based membrane technology in the form of a convenient spin column. The kit can be used to purify DNA fragments from 25 bp to 20 kb in size. The recovery rates are up to 95% in a 100 bp – 10 kb DNA fragment size range (see Fig. 1). Each GeneJET purification column has a binding capacity of up to 25 μg of DNA and can process up to 1 g of agarose gel.
The entire procedure takes just 15 min and the isolated DNA is ready to use in all common downstream applications including ligation, restriction digestion, PCR, sequencing and labeling.
Fig. 1. Recovery dependence on DNA fragment size
3
PRINCIPLE
The DNA fragment of interest is excised from an agarose gel, placed in a microcentrifuge tube, solubilized in binding buffer and applied to the column. The chaotropic agent in the binding buffer dissolves agarose, denatures proteins and promotes DNA binding to the silica membrane in the column. As an added convenience, the binding buffer contains a color indicator that allows for easy monitoring of the solution pH for optimal DNA binding. Impurities are removed with a simple wash step. Purified DNA is then eluted from the column with the elution buffer. The recovered DNA is ready for use in downstream applications.