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Title:Optimization of DNA isolation from hop cones and pellets for microsatellite analysis
Authors:ID Tacer, Matic (Author)
ID Luskar, Lucija (Author)
ID Čerenak, Andreja (Author)
ID Jakše, Jernej (Author)
ID Volk, Helena (Author)
Files:URL URL - Source URL, visit https://www.dlib.si/details/URN:NBN:SI:spr-OS4I5CDJ
 
.pdf PDF - Presentation file, download (738,84 KB)
MD5: 12E84752C369BE972738363F946D5E49
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo IHPS - Slovenian Institute of Hop Research and Brewing
Abstract:Reliable identification of hop (Humulus lupulus L.) cultivars is important for quality control and authentication in the brewing industry. DNA-based methods provide a powerful tool for this purpose, but isolation of high-quality DNA from processed hop materials such as cones and pellets can be challenging due to the presence of PCR inhibitors, including polyphenols, polysaccharides, and bitter acids. In this study we compared four cetyltrimethylammonium bromide (CTAB)-based DNA extraction protocols for hop cones and pellets, with the aim of improving yield and purity of DNA used for microsatellite genotyping. The tested methods included the standard CTAB protocol, CTAB supplemented with polyvinylpyrrolidone (PVP40), CTAB with PVP40 and activated charcoal, and CTAB with PVP10 and liquid nitrogen grinding. Additionally, a hexane pre-treatment step was evaluated with the aim to reduce the amount of PCR inhibitory compounds. DNA quality was assessed using NanoVue, Qubit, and agarose gel electrophoresis. Agarose gels showed intact high-molecular-weight DNA with minor RNA traces. Microsatellite genotyping confirmed consistent allele profiles across the first three extraction methods, thus confirming the suitability of CTAB-based methods for reliable hop genotyping.
Keywords:hop, Humulus lupulus, DNA extraction, CTAB, genotyping
Publication date:19.12.2025
Year of publishing:2025
Number of pages:str. 4-14
Numbering:Št 32
PID:20.500.12556/DiRROS-27860 New window
UDC:582.630.2
ISSN on article:2536-1988
COBISS.SI-ID:262319619 New window
Copyright:Inštitut za hmeljarstvo in pivovarstvo Slovenije
Note:Nasl. z nasl. zaslona; Opis vira z dne 19. 12. 2025;
Publication date in DiRROS:26.02.2026
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Downloads:60
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Record is a part of a journal

Title:Hmeljarski bilten
Publisher:Inštitut za hmeljarstvo in pivovarstvo Slovenije (IHPS), = Slovenian Institute of Hop Research and Brewing (IHPS)
ISSN:2536-1988
COBISS.SI-ID:287262720 New window

Document is financed by a project

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P4-0077
Name:Kmetijske rastline - genetika in sodobne tehnologije

Licences

License:CC BY 4.0, Creative Commons Attribution 4.0 International
Link:http://creativecommons.org/licenses/by/4.0/
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.
Licensing start date:01.01.2025

Secondary language

Language:Slovenian
Title:Optimizacija izolacije DNA iz storžkov in peletov hmelja za analizo mikrosatelitov
Abstract:Možnost zanesljivega določanja sorte hmelja (Humulus lupulus L.) je pomembna za zagotavljanje kakovosti in avtentičnosti v pivovarski industriji. Metode, ki temeljijo na analizi DNA, so za ta namen učinkovito orodje, vendar je izolacija visokokakovostne DNA iz procesiranih oblik hmelja, kot so storžki in peleti, zahtevna zaradi prisotnosti inhibitorjev PCR, med katerimi so polifenoli, polisaharidi ter alfa in beta kisline. V tej raziskavi smo primerjali štiri protokole za izolacijo DNA na osnovi CTAB iz hmeljnih storžkov in peletov. Naš namen je bil izboljšati koncentracijo in čistost DNA, ki jo uporabimo za genotipizacijo hmelja z mikrosateliti. Preizkušene metode so bile: standardni CTAB protokol, CTAB z dodatkom polivinilpirolidona (PVP40), CTAB z dodatkom PVP40 in aktivnega oglja ter CTAB z dodatkom PVP10 in s predhodno homogenizacijo v tekočem dušiku. Poleg tega smo preizkusili tudi predobdelavo s heksanom, katere namen je bil v izolirani DNA zmanjšati količine spojin, ki zavirajo PCR. Kakovost izolirane DNA smo določali z instrumentoma NanoVue in Qubit ter z agarozno gelsko elektroforezo s katero smo pokazali prisotnost nepoškodovane DNA visoke molekulske mase z manjšimi sledovi RNA. Genotipizacija z mikrosateliti je pokazala skladne alelne vzorce pri prvih treh metodah izolacije, kar potrjuje primernost CTAB-protokolov za zanesljivo genotipizacijo hmelja, tako iz storžkov, kot iz peletov.
Keywords:hmelj, Humulus lupulus, izolacija DNA, CTAB, genotipizacija


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