An improved method of extracting genomic DNA from preserved tissues of Capsicum annuum for PCR amplification

PCR amplifikasyonu amacıyla Capsicum annuum'un korunmuş dokularından genomik DNA edilmesinde gelişmiş bir metot

Adebayo Liasu OGUNKANMI,Bola OBOH,Bukola ONIFADE,Adeniyi Adewale OGUNJOBI,Idowu Adewumi TAIWO,Oluwatoyin Temitope OGUNDIPE

Özet

Bu calismada, farkli biber (Capsicum annuum) dokularindan genomik DNA ekstrasyonu icin bir metot sunulmustur. Yuksek miktarda polisakkarit icerdigine inanilan Capsicum orneginin korunmus dokularindan DNA ekstrasyonu icin, standart Dellaporta protokolu gozden gecirildi ve degistirildi. Kullanilan degistirilmis protokolle yuksek kalitede DNA elde edildi ve PCR ve RAPD analizleri icin uygun oldugu goruldu. Ayrica yontemin guvenilir oldugu, bazi maddelerin mevcut olmadigi durumlarda kullanilabilecegi ve fenolkloroform ekstraksiyonu gerektirmedigi anlasildi. Metot, saklama imkanlarinin bulunmadigi yerlerde bitki dokularinin birkac gun boyunca korunmasina imkan tanidi. Ayrica, doku kaynagi ne olursa olsun, iyi kalitede DNA elde edildigini tespit edilmistir. Etli mezokarp dokusundan elde edilen DNA miktari, ayni agirliktaki tohumlardan elde edilenden daha fazlaydi. Bu durum, tohumlarin sert yapisindan ve hucre iceriginin salinmasi icin hucre ceperinin tamamen parcalanamamasindan kaynaklaniyor olabilir.

Abstract

In this study we present a method for the extraction of genomic DNA from the different tissues of the Pepper (Capsicum annuum). A standard protocol of Dellaporta was reviewed and modified for DNA extraction from the preserved tissues of the Capsicum sample which was believed to contain high level of polysaccharides. The modified protocol employed yielded a high quality DNA and was found to be suitable for PCR and RAPD analyses. The procedure was also found to be reliable and suitable where some materials are not available and does not require phenol-chloroform extraction. The method also allowed for the preservation of plant tissues for some days from a locality where storage facilities are not accessible. We also discovered that irrespective of the sources of tissues, a good quality DNA was obtained. The quantity of DNA produced from the fleshy mesocarp tissue was more than the quantity obtained from the seeds of the same weight, this is probably due to the hard nature of the seeds and there may have been no complete breakdown of the cell wall to release the cellular contents.

Anahtar kelimeler: genom,dokular,Capsicum annuum,polimeraz zincir reaksiyonu,izolasyon teknikleri,izolasyon,DNA