Extraction of DNA from Virus
Mix Lysing Solution (LS) & Sodium Perchlorate (SP) in 2:1.
Mix the combined Lysing Solution & sodium Perchlorate with virus prep :18.5 ml LS+SP with 10ml virus prep (~ 10¹¹ PFU/ml ).
Incubate the lysate at 55ºC for 1hrs.
Add 30ml of phenol : [ chloroform : isoamyl alcohol (25:1)] in the proportion equal to 1:1.
Shake them by hands for 5mins.
Transfer the liquid to the centrifuge tubes, balance them & centrifuge for 15 mins at 14000 rpm at 4ºC.
Transfer the aqueous (upper) part with pipette to the clean centrifuge tubes.
Don’t worry getting protein in it.
Put the bottom layer into the waste container.
Centrifuge tubes again at 14,000 rpm for 15 min.
Now it easy to get pure liquid without protein debris.
Aqueous layer may be stored overnight at 4 ºC.
Estimate the volume of the aqueous layer and add 0.1 volume of 3M (or 5M) Sodium Acetate (pH 6.0).
Collect DNA by swirling glass rod.
Let it dry on the glass rod.
Pour off alcohol into the waste container.
Put DNA on the rod into the beaker & add 10 to 20ml of cold 76% Ethanol (-20ºC).
Let it stand for 10-20 min.
Press the glass rod with DNA against the wall of beaker to get rid of the ethanol.
Invert the stirring rod in the test tube rack and dry for 5 min.
Use 15ml plastic tubes.
Dissolve DNA in 4ml of TE.
Estimate DNA concentration using spectrophotometer  or gel to ensure DNA quantity.
Gently overlay the aqueous layer with 2 volumes of 95 % of ethanol.
