Introduction
The E.Z.N.A.® Tissue DNA Kit provides a rapid and easy method for the isolation of genomic DNA for consistent PCR and Southern analyses. Up to 30 mg tissue can be readily processed in one prep. The method can also be used for preparation of genomic DNA from mouse tail snips, cultured cells, blood, buffy coat, serum, and plasma.
Principle
E.Z.N.A.® Tissue DNA Kits use the reversible binding properties of the HiBind® matrix, a new silica-based material, combined with the speed of mini-column spin technology. A specially formulated buffer system allows genomic DNA up to 60 kb to bind to the matrix. Samples are first lysed under denaturing conditions and then applied to the HiBind® spin columns to which DNA binds, while cellular debris, hemoglobin, and other proteins are effectively washed away. High quality DNA is finally eluted in sterile de-ionized water or low salt buffer.
The use of mini-columns facilitate the binding, washing, and elution steps thus enabling multiple samples to be simultaneously processed. For more information on columns, click Flexible Multi-Format Columns.
Procedure
Mince up to 30 mg of tissue and add buffer TL. Add OB Protease (D3496& D3396) or Proteinase K (D3495&D3395) solution, and mix well for the complete lysis. Add Buffer BL, incubate at 70°C for 10 minutes and then add absolute ethanol and mix thoroughly. Allow the entire sample to pass through the HiBind® spin column and centrifuge to bund the DNA. Wash the column twice with DNA wash buffer diluted with absolute ethanol. Centrifuge the empty column at maximum speed for 2 minutes to dry the HiBind® matrix, before eluting the DNA with preheated elution buffer.
Applications
DNA purified using the E.Z.N.A.® Tissue DNA Kit is suitable for the following applications :
- PCR
- Restriction Digestion
- Southern blotting
