Can DNA Barcode Study be Done from a Museum Specimen Fixed in a Formaldehyde Solution? A Case of Emys orbicularis (Linnaeus, 1758)

Öz

DNA barcoding, a molecular taxonomy technique, has been increasingly used by herptile taxonomists in recent years. In DNA barcoding studies with museum specimens, there are difficulties in achieving success in specimens that have been exposed to formaldehyde, which is usually used as a fixative, for a long time and intensively. Here we studied the effect of formaldehyde on the application of the DNA barcode method in Emys orbicularis specimens stored in 4% formaldehyde and 70% ethanol solution since 2008 and 2014. Sanger sequence analysis of tissues taken from samples stored in both ethanol and formaldehyde solution successfully yielded sequences of 623 bp. In conclusion, the use of ethanol solutions should be preferred for mid or long-term sample storage, especially in the context of molecular studies. In cases where the use of formaldehyde is unavoidable, it may be advisable to use extremely low concentrations to increase success in molecular research.

Anahtar Kelimeler

• Cytochrome oxidase I
• European pond turtle
• DNA barcoding
• Collection
• Türkiye

Formaldehit Çözeltisindeki Bir Müze Örneğinden DNA Barkod Çalışması Yapılabilir mi? Emys orbicularis (Linnaeus, 1758) Örneği

Öz

Bir moleküler taksonomi tekniği olan DNA barkodlama, son yıllarda herptil taksonomistleri tarafından giderek daha fazla kullanılmaktadır. Müze örnekleri ile yapılan DNA barkodlama çalışmalarında, genellikle fiksatif olarak kullanılan formaldehite uzun süre ve yoğun bir şekilde maruz kalan örneklerde başarı elde etmekte zorluklar yaşanmaktadır. Bu çalışmada, herptil koleksiyon materyali olarak 2008 ve 2014 yıllarından beri %4 formaldehit ve %70 etanol çözeltisinde saklanan Emys orbicularis örneklerinde formaldehitin DNA barkod yönteminin uygulanması üzerindeki etkisi araştırılmıştır. Hem etanol hem de formaldehit çözeltisinde saklanan örneklerden alınan dokuların Sanger dizi analizi sonucunda 623 baz çiftinden oluşan dizileri başarıyla elde edilmiştir. Sonuç olarak, özellikle moleküler çalışmalar bağlamında orta ve uzun süreli numune saklama için alkol solüsyonlarının kullanılmasının tercih edilmesi gerekmektedir. Formaldehit kullanımının kaçınılmaz olduğu durumlarda ise, moleküler araştırmalarda başarıyı artırmak için son derece düşük konsantrasyonların kullanılması tavsiye edilebilir.

Anahtar Kelimeler

• Sitokrom oksidaz I
• Benekli kaplumbağa
• DNA bakrodlama
• Biyolojik koleksiyon
• Türkiye

Kaynakça

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