The E.Z.N.A.® Blood DNA Midiprep Kits are designed for the isolation of genomic DNA from up to 10 ml fresh, frozen, and anti coagulated whole blood treated with any common anticoagulant such as heparin, EDTA, or acid-citrate -dextrose. 10 ml of blood typically yields 200-250 μg genomic DNA. This method can also be used for the preparation of genomic DNA from buffy coat, serum, plasma, and cultured cells. The kit allows for the simultaneous processing of single or multiple samples in under 40 minutes.

Principle

The E.Z.N.A.® Blood DNA Midiprep Kits use the reversible binding properties of HiBind® matrix, a new silica-based material. This is combined with the speed o f midi-column spin technology. HiBind™ matrix avidly, but reversibly, binds DNA or RNA under certain optimal conditions allowing proteins and other contaminants to be removed. Nucleic acids are easily eluted with de-ionized water or low salt buffer. The use of midi-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

Red blood cells are selectively lysed and white cells collected by centrifugation. After lysis of white blood cells under denaturing conditions that inactivate DNases, genomic DNA is purified on the HiBind® Midi spin column. A specifically formulated high salt buffer system allows DNA molecules greater than 200 bases to bind to the matrix. Cellular debris and other contaminants (such as hemoglobin) are effectively washed away and high quality DNA is finally eluted in DNA Elution Buffer.

Applications

DNA purified using the E.Z.N.A.® Midiprep Blood DNA Kit is suitable for the following applications :

• PCR
• Restriction Digestion
• Southern blotting
• RT-PCR