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Pregled bibliografske jedinice broj: 311399

Interaction of Humic Acids with Human DNA: Proposed Mechanisms and Kinetics


Sutlović, Davorka; Gamulin, Stjepan; Definis-Gojanović, Marija; Gugić, Dijana; Anđelinović, Šimun
Interaction of Humic Acids with Human DNA: Proposed Mechanisms and Kinetics // Electrophoresis, 29 (2008), 7; 1467-1472 (međunarodna recenzija, članak, znanstveni)


CROSBI ID: 311399 Za ispravke kontaktirajte CROSBI podršku putem web obrasca

Naslov
Interaction of Humic Acids with Human DNA: Proposed Mechanisms and Kinetics

Autori
Sutlović, Davorka ; Gamulin, Stjepan ; Definis-Gojanović, Marija ; Gugić, Dijana ; Anđelinović, Šimun

Izvornik
Electrophoresis (0173-0835) 29 (2008), 7; 1467-1472

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
humic acid; humic acid intercalation; PCR inhibition mechanisms; Quantitative Real Time PCR; DNA quantification

Sažetak
Human DNA quantification by Quantitative Real Time PCR (QRT-PCR) has gained great importance in forensic DNA and ancient DNA studies. However, in such samples, DNA quantification is impaired by frequently present humic acid (HA). We have previously shown that the addition of synthetic HA inhibits QRT-PCR. In this study we investigated the possible mechanisms of HA interaction with human DNA, and kinetics of QRT-PCR inhibition. In QRT-PCR with pure human DNA and no HA added, VMAX was 40. With DNA sample containing 4  g/ml of HA, VMAX was 30.30 while the addition of extra Taq polymerase to the same sample changed VMAX into 38.91, amplifying between 80 and 90% of input DNA. The KM/VMAX ratio in all the samples remained constant, indicating that the mechanism of HA inhibition of QRT-PCR is uncompetitive by nature. Moreover HA shifts the human DNA melting temperature point (Tm) from 75oC to 87oC and inhibits DNase I mediated DNA cleavage, most probably affecting the enzyme’ s activity.

Izvorni jezik
Engleski

Znanstvena područja
Kemija, Biologija, Temeljne medicinske znanosti



POVEZANOST RADA


Projekt / tema
141-2160800-0333 - DNA analiza ranosrednjovjekovne populacije s područja južne Hrvatske (Šimun Anđelinović, )
216-2160800-0799 - Antropološka analiza kostura ranosrednjovjekovne populacije iz južne hrvatske (Marija Definis-Gojanović, )
216-2160800-0655 - Metali u kosturima iz starohrvatskih grobova u Kliškoj i Bribirskoj županiji (Davorka Sutlović, )

Ustanove
KBC Split,
Medicinski fakultet, Split

Citiraj ovu publikaciju

Sutlović, Davorka; Gamulin, Stjepan; Definis-Gojanović, Marija; Gugić, Dijana; Anđelinović, Šimun
Interaction of Humic Acids with Human DNA: Proposed Mechanisms and Kinetics // Electrophoresis, 29 (2008), 7; 1467-1472 (međunarodna recenzija, članak, znanstveni)
Sutlović, D., Gamulin, S., Definis-Gojanović, M., Gugić, D. & Anđelinović, Š. (2008) Interaction of Humic Acids with Human DNA: Proposed Mechanisms and Kinetics. Electrophoresis, 29 (7), 1467-1472.
@article{article, year = {2008}, pages = {1467-1472}, keywords = {humic acid, humic acid intercalation, PCR inhibition mechanisms, Quantitative Real Time PCR, DNA quantification}, journal = {Electrophoresis}, volume = {29}, number = {7}, issn = {0173-0835}, title = {Interaction of Humic Acids with Human DNA: Proposed Mechanisms and Kinetics}, keyword = {humic acid, humic acid intercalation, PCR inhibition mechanisms, Quantitative Real Time PCR, DNA quantification} }

Časopis indeksira:


  • Current Contents Connect (CCC)
  • Web of Science Core Collection (WoSCC)
    • Science Citation Index Expanded (SCI-EXP)
    • SCI-EXP, SSCI i/ili A&HCI
  • Scopus
  • MEDLINE





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