Rezumatul tezei de doctorat - USAMV Cluj-Napoca

So, DNA purities obtained after kit extraction (Qiagen) varied between 1,02 and 1,51,
with an average of 1,250, at 260/280 nm, and the DNA quantity obtained was between 2,54
and 14,00 ng/µl, with an average of 5,543 ng/µl. Analyzing the DNA purities after the
extraction with PBS solution, we observate that the samples have values between 1,10 and
1,51, with an average of 1,394, at 260/280 nm, and DNA quantities varied between 5,23
ng/µl and 86,56 ng/µl, with an average of 30,653 ng/µl.
II.5. THE DNA AMPLIFICATION RESULTS OF FUNGAL SPECIES
FROM SAPROLEGNIACEAE FAMILY
II.5.1. DNA amplification using PCR
The amplification reactions were individually performed, in a final volume of 25 µl,
with a thermocycler Eppendorf. In a preliminary phase, the amplification process was
applied on 20 samples, 10 of the fungal DNA being extracted with kits (QIAGEN) and 10
extracted with PBS solution. This test was done to observe which of the two methods used,
realizes a more precisely DNA amplification, without any unspecific product.
The second protocol used, described in material and method, didn’t give the expected
results at fungi, so we abandoned it. In the figure 3, we present the comparative results
regarding the electrophoretic profiles of the fragments amplified with the two primer pairs.
Because we used in the DNA extractions, PBS solution and QIAGEN kit, and the
universal primers, the electrophoresis results convinced me to continue with the DNA kit
extraction (QIAGEN), and ITS1 - ITS4 primer pair.
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