MEDLABORATORIJSKA PRIMERJAVA REZULTATOV GENOTIPIZACIJE FIGE Z MIKROSATELITSKIMI MARKERJI (Ficus carica L.) IN DOLOČITEV REFERENČNIH ALELOV

Matjaž HLADNIK, Jernej JAKŠE, Bouchaib KHADARI, Sylvain SANTONI, Dunja BANDELJ

Povzetek


Mikrosateliti so se izkazali kot zelo uporabni markerji pri genetskih raziskavah rastlin. Zaradi odstopanj dolžin enakih alelov v različnih laboratorijih, je primerjava rezultatov med laboratoriji pogosto težavna. Namen raziskave je bil primerjati rezultate genotipizacije dveh laboratorijev, ovrednotiti genetske parametre mikrosatelitskih markerjev in določiti dolžine referenčnih alelov za sorte fig istrskega polotoka.

Rezultati genotipizacije devetdesetih vzorcev fige (Ficus carica L.) so bili primerljivi med laboratorijema, kljub razlikam, ki smo jih opazili pri primerjavi elektroferogramov različnih sistemov kapilarnih elektroforez. Razlike med dolžinami enakih alelov med laboratorijema so bile odkrite zaradi različnih metod PCR in analitske opreme, vendar pa so bile razlike med aleli istega lokusa ohranjene. Pri lokusu FSYC01 smo ugotovili izpad alela, kar je privedlo do napačne identifikacije; namesto 28 heterozigotov smo posameznike določili kot homozigote. Ugotovljena lastnost nakazuje na nezanesljivost lokusa FSYC01. Izpad alela smo pripisali uporabi ekonomične metode PCR ali uporabi protokola PCR s postopnim nižanjem temperature prileganja začetnih oligonukleotidov.

Genotipi štiriindvajsetih referenčnih sort istrskega polotoka so bili potrjeni v obeh laboratorijih. Rezultati raziskave bodo prispevali k uporabi bolj zanesljivih mikrosatelitskih markerjev, z večjo močjo razlikovanja in posledično k zanesljivi standardizaciji rezultatov z drugimi genetskimi raziskavami fige.

Ključne besede


mikrosatelitski marker; referenčni genotip; medlaboratorijska primerjava; Ficus carica L.

Celotno besedilo:

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Literatura


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DOI: http://dx.doi.org/10.14720/aas.2018.111.1.14

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