Rezumatul tezei de doctorat - USAMV Cluj-Napoca

The DNA rapid extraction using PBS solution, applied on 69 samples, performed
in the following way: there was weighted 120 mg of fungal tissue in an Eppendorf tube and
added 200 µl PBS solution. The samples were centrifuged at 3000 rpm, for 20 minutes, then
the supernatant was removed. This operation repeated five times, until the fungal DNA
pellet was clean. Then, the samples were introduced in a marine bath, at 95°C for 15
minutes, to disrupt the cell wall. The samples were dried for 30 minutes, and then was addes
TE (Tris-EDTA) solution for DNA rehydrating.
For the DNA rapid extraction method with NE solution are used some chemical
substances, the solutions being made in a laboratory. The materials and expendables were
the following: Eppendorf pipettes, with adjustable volumes; Eppendorf tubes; pipette tips of
different dimensions (large, medium and small); sterile water.
I.3.6. DNA quantification using direct method of DNA purity and concentration with
Nanodrop ND-1000 spectrophotometer
For the establishing of DNA purity and concentration, there was used Nanodrop ND-
1000 spectrophotometer. To be able to find the DNA concentration extracted from fungal
samples, we used the direct method, which involves the optical density measurement of
DNA sample, at 260/280 nm ripple length on a spectrophotometer, in UV/VIS domain.
I.3.7. ITS region from fungi and primers used
At the fungi and other eukaryotes, there are two locations for rDNA: the nuclear and
the mitochondrial genome. The last one contains two genes which are coding the small and
large mitochondrial genes of rDNA. Fungal nuclear rDNA is generally structured in a
tandem repeated nuclear unit. A rDNA unit, illustrated in fig.2, includes 3 rRNA genes:
small nuclear (18S) rRNA, 5,8S rRNA and large nuclear (28S) rRNA gene. In a unit, the
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