实验概要
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 analysis. Up to 30 mg tissue or up to 1 cm sections of mouse tail can be readily processed in one time. The method can also be used for preparation of genomic DNA from mouse tail snips, blood, buffy coat, serum, and plasma. The kit allows single or multiple, simultaneous processing of samples. There is no need for phenol/chloroform extractions, and timeconsuming steps such as precipitation with isopropanol or ethanol, are eliminated. DNA purified using the E.Z.N.A.® Tissue DNA method is ready for applications such as PCR*, Southern blotting, and restriction digestion.
Carry out disruption, homogenization, Protease digestion, and loading onto HiBind® DNA column as indicated previous protocols. Instead of continuing with centrifugation, follow steps blow.
实验步骤
Note: Please read through previous section of this book before using this protocol.
1. Prepare the vacuum manifold according to manufacturer’s instruction and connect the HiBind® DNA V-Spin column to the manifold.
2. Load the sample into HiBind® DNA V-spin column.
3. Switch on vacuum source to draw the sample through the column and turn off the vacuum.
4. Wash the column by adding 500 ul Buffer HB, draw the wash buffer through the column by turning on the vacuum source.
5. Wash the column by adding 700 ul DNA wash buffer, draw the wash buffer through the column by turning on the vacuum source.
6. Wash the column again by adding 700 u DNA wash buffer, draw the wash buffer through the column by turning on the vacuum source.
7. Assemble the column into a 2 ml collection tube and transfer the column to a micro centrifuge. Spin at maxi speed (no more than 20,000 x g) for 2 minute to dry the column.
8. Place the column in a clean 1.5 ml microcentrifuge tube and add 50-100ul DNA elution buffer. Stand for 1-2 minute and centrifuge 1 minute to elute DNA.