Digitalni repozitorij raziskovalnih organizacij Slovenije

Iskanje po repozitoriju
A+ | A- | Pomoč | SLO | ENG

Iskalni niz: išči po
išči po
išči po
išči po

Možnosti:
  Ponastavi


Iskalni niz: "ključne besede" (PCR) .

1 - 7 / 7
Na začetekNa prejšnjo stran1Na naslednjo stranNa konec
1.
The influence of storage conditions and DNA extraction protocol on the results of molecular analysis of the European spruce bark beetle (Ips typographus L.)
Zina Devetak, Andreja Kavčič, Maarten De Groot, Barbara Piškur, 2023, izvirni znanstveni članek

Povzetek: One of the key steps of the molecular identification of bark beetles is obtaining a sufficient quantity of high-quality DNA extract. In this study, we investigated the influence of different storage procedures for Ips typographus (L.) specimens and various DNA extraction protocols on the quantity and quality of DNA intended for use in molecular diagnostics. Adult beetles were frozen at -20 °C, either dry or in ethanol. We tested four different protocols for DNA extraction. We compared the quantity of extracted DNA and assessed its quality with PCR and Sanger sequencing. Different storage protocols had no significant effect on the quantity of DNA extracted. However, freezing specimens in ethanol provided higher-quality DNA for molecular applications. Only two of the extraction protocols produced sequenceable amplicons, and the difference in the amount of extracted DNA between them was not significant. We propose the optimal combination of storing specimens in ethanol at -20°C and using the Nucleospin Insect DNA extraction kit from Macherey Nagel, enabling a timeefficient identification process.
Ključne besede: early detection, specimen storage, total DNA extraction, PCR, polymerase chain reaction, Sanger sequencing, molecular diagnostics
Objavljeno v DiRROS: 02.02.2024; Ogledov: 328; Prenosov: 101
.pdf Celotno besedilo (1,17 MB)
Gradivo ima več datotek! Več...

2.
Verifikacijsko poročilo - LVG POS 021
Zina Devetak, 2023, elaborat, predštudija, študija

Ključne besede: Ceratocystis platani, platanov obarvani rak, diagnostični protokol, molekularna analiza, ekstrakcija DNA, PCR v realnem času, verifikacija
Objavljeno v DiRROS: 07.06.2023; Ogledov: 288; Prenosov: 0

3.
Verifikacijsko poročilo - LVG POS 020
Zina Devetak, 2023, elaborat, predštudija, študija

Ključne besede: Geosmithia morbida, bolezen tisočerih rakov, diagnostični protokol, molekularna analiza, ekstrakcija DNA, PCR v realnem času, verifikacija
Objavljeno v DiRROS: 07.06.2023; Ogledov: 301; Prenosov: 0

4.
5.
PCR primers comparisons for a successful Tuber spp. DNA region amplification in routine identifications
Tina Unuk Nahberger, Hojka Kraigher, Tine Grebenc, 2020, izvirni znanstveni članek

Povzetek: Since late 20th century DNA sequencing became the method of choice method in precision species identification. The ITS region is one of the official fungal barcoding DNA markers, although in some cases sequencing of the ITS re-gion may, due to misidentification, mislabeling or nomen-clature errors in public databases, lead to incorrect or insuf-ficient identification, as is currently a case in the genus Tu b e r. The aim of this study was to test, which ITS primer pairs are most appropriate and optimal for Tu b e r species DNA region amplification. Thereby we (1) compared ampli-fication success for different Tu b e r species using fungal spe-cific primer pair ITS1f and ITS4 and (2) compared amplifi-cation success using different ITS primer pair combinations in amplifying DNA region an example species Tuber aesti-vum. Based on results, Tuber aestivum was one of the most reluctant Tu b e r species in this study and in most cases failed to amplify with the above primer pair. After comparing dif-ferent ITS primer pairs, we conclude that the primer pair ITS5 and ITS7 is the most appropriate primer pair for ampli-fication DNA region of T. ae stiv um as it resulted in high am-plification success from ectomycorrhizal root tips. Based on sequences, gained from public databases, we found that ITS1f and ITS6 primers have a mismatch in one base pair compared to the target sequence of Tuber aestivum, thus re-sulting in poor or no amplification success. Although prim-er pair ITS5 and ITS7 in our study was proven to be the most appropriate primer pair in amplifying DNA region Tu b e r aestivum species, further analysis about appropriateness of it for a general barcoding and identification of ectomycorrhiza in complex community samples is needed.
Ključne besede: Tuber spp., ITS region, PCR amplification, ITS primers
Objavljeno v DiRROS: 30.07.2020; Ogledov: 1577; Prenosov: 1165
.pdf Celotno besedilo (6,81 MB)
Gradivo ima več datotek! Več...

6.
7.
Vpliv glivnih in rastlinskih sekundarnih metabolitov na verižno reakcijo s polimerazo (PCR)
Nejc Thaler, Marko Bajc, 2013, pregledni znanstveni članek

Povzetek: Sekundarni metaboliti so organske spojine, ki jih najdemo pri glivah in rastlinah, kjer imajo vlogo obrambnih in signalnih molekul ali zagotavljajo druge selekcijske prednosti, niso pa neposredno vpleteni v rast, razvoj in razmnoževanje organizma. Pri delu s tehnikami DNA so pogosto ravno sekundarni metaboliti tisti, ki posredno ali neposredno vplivajo na uspešnost verižne reakcije s polimerazo (PCR) ali reverezno transkriptazo, in sicer tako, da otežujejo celično lizo, povzročajo razpad nukleinskih kislin ali neposredno ovirajo delovanje encima polimeraze pri pomnoževanju tarčne DNK. Glavna ovira pri aplikaciji tehnike PCR v rutinski diagnostiki je priprava visoko kvalitetne DNA brez inhibitorjev. Še posebej to velja pri izolaciji DNA iz lesnatih rastlin (Minafra in sod., 1992) in vzorcev tal (Tsai in Olson, 1991). Večina standardnih postopkov izolacije nukleinskih kislin ne odstrani rastlinskih polisaharidov in polifenolnih komponent, ki imajo lahko neposreden vpliv na pomnoževanje s PCR (Demeke in Adams, 1992). Poskusi za premostitev tovrstnih ovir vključujejo bolj dovršene metode za izolacijo nukleinskih kislin in PCR, ki vključujejo uporabo pospeševalcev PCR za odstranitev ali zmanjšanje vpliva inhibitorjev PCR. Ta pregled je osredotočen na pristope za odstranitev ali zmanjšanje vplivov rastlinskih in glivnih sekundarnih metabolitov iz vzorcev tal, različnih rastlinskih tkiv in razkrojenega lesa zaradi pomena tovrstnih raziskav za gozdarstvo in lesarstvo.
Ključne besede: sekundarni metaboliti, izolacija DNA, pomnoževanje DNA, verižna reakcija, polimeraza, PCR
Objavljeno v DiRROS: 12.07.2017; Ogledov: 4916; Prenosov: 2262
.pdf Celotno besedilo (685,41 KB)

Iskanje izvedeno v 0.23 sek.
Na vrh