Genomic DNA from Tissue

-Dominik Haudenschild from Red Book January 2001

• Crush Tissue to fine powder with mortar & pestle
• Add 1.2ml Digestion Buffer per 100mg tissue
• Digest overnight at 50°C with gentle rocking
• Add 1 volume Phenol/Chloroform/Isoamyl
• Centrifuge 10minutes in swinging bucket rotor at 10,000g
-if phases not resolved, re-extract with 1 volume digestion buffer
-if white interface is thick, re-extract with 1 volume Phenol/Chlo/Isoam
a. if you need smaller DNA
-remove aqeuous layer and add 1/2 volume 7.5M Ammonium Acetate
-precipitate with 2 volumes 100% EtOH
b. If large size intact genomic DNA is needed
-dialyze against large volume of TE twice