I need to extract DNA from spores from herbaria material. I've used the Phenol-Chloroform method with success, but I'm interested in a Kit.
I had great results with the DNEasy® Blood and Tissue Kit (Qiagen). Using a bead-beating step to break down the cell wall (would fill a 1.5uL microfuge tube with beads to the meniscus of the sample then vortex for 2 minutes (I do not remember the exact size, but I think 2um was the bead size. Then added enzymatic lysis buffer before proceeding with the rest of the protocol.
E.Z.N.A.® High Performance (HP) Fungal DNA Kit worked good for herbarium material. It´s based on a chloroform extraction. I think for old DNA samples the advantage of kits is (beside time saving) the separation of highly fragmented DNA (< 200 bp) from fragments of higher mol. weight. The small fragments might otherwise "catch" away your primers without being suitable for PCR (because they are simply not long enough to reach from one primer binding site to the other).
I agree with Mark. I have used it for Aspergillus spp. it was Excellent
We use fungal Genomic DNA Kit from Zymo Research for Aspergillus niger.
but I assume you have cultures... I want to extract from spores.
We also use DNeasy Plant Mini Kit (Qiagen), but haven't tried it on spores.
Hi Eduardo!
We use FTA Cards (GE Healthcare Life Sciences) to extract spores DNA from herbaria material. We are managing to extract DNA from very old type specimens (with over one hundred years)!
you can check out this webpage: www.thermofisher.com/microbiome describing microbiome tools and workflows
You can use Promega A1120, it is really good with fungi. It can give you high quantity and quality of G-DNA.
Hi Eduardo
You can follow the procedure described in the chapter with link below. It works for isolating DNA from fungal spores also.
Chapter Heterologous Expression of Fungal Secondary Metabolite Pathw...
Thanks
Mohit
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