Description

The EXTRACTME DNA GEL-OUT kit is designed for the rapid and efficient purification of DNA fragments directly from agarose gels (standard and low-melting point agarose gels run in either a TAE or TBE buffer). Agarose, ethidium bromide and other contaminants from a sample are effectively removed in the purification process. The kit enables the purification of DNA fragments from 50 bp to 30 kb, as well as plasmid and genomic DNA. However, purification of fragments smaller than 100 bp and larger than 10 kb will result in decreased recovery rates. The purified DNA can be used in common downstream applications.

The purification protocol and buffer formulations were optimised for high yields and DNA purity.

The DNA purification procedure utilises spin minicolumns with membranes which efficiently and selectively bind nucleic acids. In the first step, the DNA fragment is excised from an agarose gel and incubated in the GB Buffer, which enables gel fragment solubilisation and protein degradation. As an added convenience, the binding buffer contains a colour indicator, which facilitates easy monitoring of the solution’s pH for optimal DNA binding. The two-step washing stage efficiently removes impurities and enzyme inhibitors. The purified DNA is eluted using either a low ionic strength buffer (Elution Buffer) or water (pH 7.0-9.0) and can be used directly in all downstream applications such as PCR, Southern blotting, DNA sequencing, enzymatic restriction, ligation and so forth or stored until ready to use.

The quality of each production batch (LOT) of the EXTRACTME DNA GEL-OUT kit is tested using standard QC procedures. The purified DNA concentration, quality and stability are evaluated by gel electrophoresis and spectrophotometer. In addition, the functional quality is tested by qPCR and digestion with restriction enzymes.