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

Year 2018, Volume: 44 Issue: 2, 135 - 148, 11.10.2018

Murat Güler , Filiz Tokgöz Güler Ertan Mahir Korkmaz , Mahir Budak

Abstract

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.

 

Keywords

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

Abstract

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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