Year 2020, Volume 48, Issue 4, 333 - 339, 06.07.2020

Zübeyde UĞURLU AYDIN Mustafa K. ŞENOVA Marcus KOCH Ali DÖNMEZ

https://doi.org/10.15671/hjbc.663505

Abstract

DNA from Leaf or Stem: A Comparative Work on Dianthus L. for DNA Barcoding Analysis with Four Commercial Extraction Kits

Abstract

GeneAll GenExTM Plant kiti, GeneMATRIX Plant & Fungi DNA Purification kiti, DNeasy Plant kiti ve E.Z.N.A. Plant DNA kiti yaprak ve gövde dokusu DNA barkodlama çalışmaları için kaliteli ve yüksek genomik DNA elde etmek amacıyla test edilmiştir. Geleneksel olarak yaprak dokusu genomik DNA kaynağı olarak kullanılmaktadır fakat Dianthus taksonları yaprak sklerankima dokusu nedeniyle bu bakımdan sorunludur. Genomik DNA, üretici firmanın sağladığı protokole göre elde edilmiştir ve yedi barkod bölgesi PCR yöntemiyle çoğaltılmıştır. DNA konsantrasyonu florometrik yöntemle ölçülmüş, PCR ürünleri agaroz jelde görüntülenmiştir. Sonuçlar, DNA barkodlama çalışmalarında kullanılabilecek PCR başarısı bakımından değerlendirilmiştir. Ayrıca, kullanılan ekstraksiyon kitleri zahmet, fiyat ve zaman bakımından değerlendirilmiştir. Sonuçlarımıza göre, yaprak ve gövde dokusundan genomik DNA ekstraksiyonu başarıyla gerçekleştirilmiştir. Tüm ekstraksiyon kitleri PCR amplifikasyonu bakımından başarılı olmasına rağmen en yüksek verim GeneMATRIX kiti kullanılarak elde edilmiştir.

Keywords

Dianthus, DNA barcoding, DNA isolation, PCR success, stem

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