The RecoverEase DNA isolation kit quickly isolates high-molecular-weight genomic DNA from a variety of tissues without organic solvent extractions or ethanol precipitation (Figure 1). The procedure involves physical disaggregation of the tissue, coarse filtration, and isolation of the cell nuclei via short centrifugation, followed by incubation with a highly active protease solution containing proteinase K to predigest the cellular proteins. Digestion continues as the DNA preparation is transferred to a convenient, free-floating dialysis cup that allows overnight dialysis of the digested peptides across a semipermeable membrane. After dialysis, the fully hydrated, purified high-molecular-weight DNA is simply collected from the dialysis cup. The resulting DNA, which consistently measures from 100 to 500 kb in length, is ready for immediate use in experimental applications including Southern blot analysis, the polymerase chain reaction (PCR), analysis by pulsed-field electrophoresis, or the recovery of lambda shuttle vectors from transgenic animals.

Figure 1: Recoverease Method vs. Phenol-Chloroform Method of DNA Isolation

Shows Recoverease method takes takes only 17 hours as compated to 3 days for organic extraction.

The benefits of the RecoverEase DNA isolation kit include the early removal of nucleases in the cell cytosol, elimination of toxic organic solvents, such as phenol and chloroform, and the reduction of shearing forces associated with extractions and excessive handling of the DNA. In addition, the DNA is fully hydrated and ready for immediate use after dialysis.

Certain tissues, like bladder and skin, are not amenable to homogenization or nuclei isolation and are thus not recommended starting materials for genomic DNA isolation using the RecoverEase DNA isolation kit.

For Research Use Only. Not for use in diagnostic procedures