Murat GÜLER, Filiz TOKGÖZ GÜLER, Ertan Mahir KORKMAZ, Mahir BUDAK

Böcek Dokularından DNA İzolasyonu Yöntemlerinin Kalite, Verim ve Maliyet Açısından Karşılaştırılması

Günümüzde moleküler biyoloji ve biyoinformatik alanındaki gelişmelere paralel olarak metagenomik, biyoteknoloji, tüm genom dizilemesi, genom evrimi gibi alanlarda çok sayıda araştırma yapılmaktadır. Böcek türleri ile yapılan çalışmalarda ilk ve en önemli adım etkili bir DNA izolasyonu yönteminin kullanılmasıdır. Kitin yapılı bir dış iskelete sahip olan böceklerin çoğunun küçük canlılar olması DNA izolasyonunu zorlaştırmaktadır. Yapılan bu çalışma ile Cephus pygmeus (Linnaeus, 1767) türüne ait bireylerden dokuz farklı DNA izolasyon yöntemi ile total DNA izolasyonu gerçekleştirilmiştir. Elde edilen DNA örnekleri verim, saflık, maliyet, zaman ve PCR başarısı kriterleri altında değerlendirilmiştir. Maliyet ve kullanılan kimyasalların toksisitesi göz önüne alındığında ise tuzla çöktürme yönteminin böcek dokularından DNA izolasyonu için en uygun yöntem olduğu yorumlanmıştır.

Anahtar Kelimeler:

PCR, DNA İzolasyon Yöntemleri, Böcek Genomik DNA’sı, DNA Kalitesi, Cephus pygmeus

Comparison of DNA Isolation Methods From Insect Tissues in Terms of Quality, Yield and Cost

Today, there are many studies conducted in the areas such as metagenomics, biotechnology, whole genome sequencing, genome evolution, and so on parallel to the developments in molecular biology and bioinformatics in molecular biology and bioinformatics. The first and most important step in studies with insect species is the use of an effective DNA isolation method. Small body and external chitin skeleton of the insects cause difficulties in isolating the insect DNAs. In this study, nine different DNA isolation methods were applied to carry out total DNA isolation from the individuals belonging to the species Cephus pygmeus (Linnaeus, 1767). The results were interpreted in terms of DNA yield, purity, cost, time and PCR performance criteria. When the cost and the toxicity of the used chemicals are taken into consideration, it is interpreted that the salting out method is the most suitable method for DNA isolation from insect tissues.

Keywords:

DNA Isolation Protocols, Insect Genomic DNA, DNA Quality, PCR, Cephus pygmeus,

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