The E.Z.N.A.® HP Tissue DNA Maxi Kit is designed for the efficient recovery of genomic DNA up to 60 kb in size from tissue samples up to 2 grams in size. The specially formulated buffer system ensures the optimal lysis of tissues rich in fat, polysaccharides, and fibers such as brain, adipose, and muscle. This kit can also isolate DNA from molluscs, insects, arthropods, roundworms, flatworms, and other invertebrate tissue samples rich in mucopolysaccharides. The procedure relies on the well established properties of the cationic detergent, cetyltrimethyl ammonium bromide (CTAB), in conjunction with the selective DNA binding of Omega Bio-tek’s HiBind® matrix.
Samples are homogenized and lysed in a high salt buffer containing CTAB and digested with proteinase. After addition of chloroform, the homogenate is separate into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted and BL Buffer is added to provide appropriate binding conditions. The sample is then transferred to the HiBind® DNA Maxi Column, where the genomic DNA binds to the membrane and salt and other contaminants are efficiently washed way. High-quality genomic DNA is eluted with Elution Buffer or water. Purified DNA is suitable for most downstream applications such as endonuclease digestion, thermal cycle amplification, and hybridization techniques.
Each HiBind® DNA Maxi column can bind approximately 2.5 mg genomic DNA. Using greater than 2 g tissue or 5 x 108 cells is not recommended.
Genomic DNA purified using the E.Z.N.A.® HP Tissue DNA Midi and Maxi Kit. Purified genomic DNA from various animal tissues were isolated with the E.Z.N.A.® HP Tissue DNA Midi and Maxi Kits. DNA was extracted from animal tissues as follows: lane 1 from 1 g pig brain, lane 2 from 200 mg mouse liver, lane 3 from 300 mg fish liver, lane 4 from 200 mg chicken spleen, lane 5 from 500 mg pig muscle, lane 6 from 200 mg chicken liver, lane 7 from 500 mg frog muscle, and lane 8 from 300 mg chicken heart. The 3% of each eluate genomic DNA was analyzed on a 0.8% agarose gel to demonstrate yield and quality of the DNA. M: Lambda Hind III DNA Marker.