Comparison of DNA extraction protocols for PCR-based techniques in wheat

Abstract

Molecular biology and genetic studies requires high quality DNA isolated from plants. Therefore, isolation of pure and high quality DNA in a short time is important. However, since different plants have different chemical composition, in even close species need specific genomic DNA isolation protocol. There are a large number of protocols for wheat. Therefore, which method is more applicable, safer and easier should be determined. For this purpose three DNA extraction protocols were used. The DNA samples extracted from these protocols were compared for their purity and concentrations. The quality and amount of DNA obtained was determined by NanoDrop Spectrophotometer (A260/280 absorbance) and agarose gel electrophoresis. Additionally DNAs extracted were amplified by polymerase chain reactions (PCR) and the suitability of isolated DNAs for PCR reactions were also determined. The results show that with the CTAB DNA extraction methods were extracted the highest DNA concentrations in wheat (Triticum avestivum L.) compared to other protocols.

Keywords

• DNA isolation,wheat,PCR,SSR

Buğdayda PCR'a dayalı teknikler için DNA ekstraksiyon protokollerinin karşılaştırılması

Öz

Moleküler biyoloji ve genetik çalışmalar, bitkilerden izole edilmiş yüksek kaliteli DNA gerektirir. Bu nedenle, saf ve yüksek kalitede DNA'nın kısa sürede izolasyonu önemlidir. Bununla birlikte, farklı bitkiler farklı kimyasal bileşime sahip olduklarından, yakın türlerde bile spesifik genomik DNA izolasyon protokolüne ihtiyaç vardır. Buğday için çok sayıda protokol vardır. Bu nedenle, hangi yöntemin daha uygulanabilir, daha güvenli ve daha kolay olduğu belirlenmelidir. Bu amaçla üç DNA ekstraksiyonu protokolü kullanılmıştır. Bu protokollerden elde edilen DNA örneklerinin saflıkları ve konsantrasyonları karşılaştırılmıştır. Elde edilen DNA'nın kalitesi ve miktarı NanoDrop Spektrofotometre (A260 / 280 absorbans) ve agaroz jel elektroforezi ile belirlenmiştir. Ayrıca, ekstrakte edilen DNA'lar polimeraz zincir reaksiyonları (PCR) ile amplifiye edilmiş ve izole edilmiş DNA'ların  PCR reaksiyonları için uygunluğu da belirlenmiştir. Sonuçlar, CTAB DNA ekstraksiyon yöntemiyle diğer protokollere kıyasla buğdayda (Triticum avestivum L.) en yüksek DNA konsantrasyonlarının elde edildiğini göstermiştir.

Anahtar Kelimeler

• DNAizolasyonu,buğday,PCR,SSR

Kaynakça

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