Hello Perumal Sasidharan,

I'm presuming you did not get fungal DNA or sufficient amounts of DNA after extraction method? Can I ask what protocol you followed? There are several rapid extraction methods available. I have personally used a rapid method developed by Don Liu et al and i have attached for you to try out.

I use approx 5 mm diameter circular section of mycelia to start with. Some pitfalls would include removal of supernatant from DNA pellet prior to wash step which can be very faint to practically invisible in some cases! Also allow adequate time to air dry ethanol before resuspending in buffer.

Good luck!

Hello Perumal,

The method given to you by Katrina is correct.

I have also a second method, which has worked just fine for me. Add 1% of β-2-mercaptoethanol to any lysis buffer that you use. Subsequently, add your sample and incubate at 55 °C for 3 h. Then you need to homogenize the sample by any means at your disposal. After this point you can start using your DNA-extraction method.

For best result I would suggest to include a single freeze/thaw cycle consisting of freezing at −80 °C overnight and thawing at 95 °C for 15 min the next day.

I have also included my own publication where I validated and used this method.

Good luck,

Ali

ADD IIQUID NITROGEN TO FUNGI

GRIND IT 

TAKE POWDER AND EXTRACT DNA BY CLOROPH-PHENOL METHOD

LAST TIME WE GOT  GOOD RESULT

Bro ask Nataraj sirz lab, they have done great, contact ragavan, some tricks n tips u have missed bro