Rajesh, we used such method http://www.biotechniques.com/multimedia/archive/00010/96216bm14_10362a.pdf

with small modifications. NaAc is essential for DNA purification.

This method was good too:

http://www.sclerotia.org/lab/sclerotinia-dna-miniprep

you can use the kits of FavorPrep Genomic DNA Mini Kit (Blood/Cultured Cell) from favorgen as in the attach

you need to perform the preparation steps as follow:

i. Harvest appropriate number of fungus cell (up to 5 x ) to a 1.5ml

microcentrifuge tube (not provided) and centrifuge at 5,000 x g

for 10 minute. Then discard the supernatant.

ii. Add 600 μl of sorbitol buffer (1.2 M sorbitol; 10 mM CaCl ;

0.1 M Tris-HCl pH 7.5; 35mM ß-mercaptoethanol) and resuspend the pellet.

iii. Add 200 U of lyticase or zymolase. Incubate for 30 minutes at 30ºC.

iv. Centrifuge the mixture at 2,000 x g for 10 minutes to harvest the spheroplast,

and then remove the supernatant.