Asmalarda Vitis vinifera L. Farklı Dokulardan izole Edilen DNA'ların RAPD Random Amplified Polymorphic DNA Tekniğinde Kullanımı Üzerinde Bir Araştırma

Öz

Asmalar ı n vejetatif otsu dokular ı sürgün ucu-genç yapraklar, sülükler ve bir ya şl ı dalları ndan DNA izolasyonu için kullan ı lan, s ı ras ı yla Lodhi ve ark. 1994 ve Lin ve Walker 1997 taraf ı ndan geliştirilen yöntemlerin, dokular ı n farkl ı yetiştirme koşullar ı ndan bağ , sera, doku kültürü bitkileri al ı nmas ı durumunda DNA verimlilikleri ve RAPD tekni ğinde kullan ı labilirlikleri incelenmi ştir. Lodhi ve ark. 1994 'na ait yöntemde, ba ğdan al ı nan ancak bekletildikten sonra kullan ı lan sürgün ucu-genç yapraklar örnekleri ile, sülüklerin kullan ı ldiğı durumlarda 10/4 DNA/g doku düzeyinden daha az yetersiz DNA elde edilirken; yetersiz düzeyde olmamakla beraber, doku kültürü ve sera bitkilerinden de 20-53 ,ug DNA/g doku düzeyinde düşük DNA miktar ı elde edilmiştir. Yöntemde, so ğ uk ethanol ilavesi ve 4°C'de DNA iplikçiklerinin olu şmad ığı aşamadan sonra gerçekleştirilen modifı kasyon sonucunda, her dört koşulda da tatmin edici DNA miktarlar ı na ulaşı lm ışt ı r. Bir yaşl ı dallardan Lin ve Walker 1997 'a göre izole edilen DNA miktarlar ı 28-40 ,ug DNA/g doku araşt ı r ı c ı lar ı n bulguları ile uyumlu bulunmuştur.

Anahtar Kelimeler

• Vitis vinifera L.
• DNA izolasyonu
• RAPD

A Research on the RAPD Assay of DNA lsolation from Different Tissues of Grapevine Vitis vinifera L.

Abstract

The DNA yields of two grape DNA isolation methods that of Lodhi et al. 1994 and Lin and Walker 1997 which are suitable for young vegatative tissues and one year old canes respectively, were determined in the tissue were obtained from different growing conditions vineyard, tissue culture and greenhouse plants . Isolated DNA was also tested for RAPD procedure. Insufficient DNA quantity which is less than 10 ,ug DNA/g tissue was obtained when the stored shoot tips-young leaves tissue were used for DNA isolation in Lodhi et al. 1994 method. Also, small quantities of DNA, 20-53 pg DNA/g tissue were extracted from the shoot tips-young leaves of tissue culture and greenhouse plants. Therefore, a few modification was carried out in the protocol begining from the stage of DNA was not visible in ice-cold ethanol. Our modification of Lodhi et al. 1994 method produced good DNA yield in every conditions. The amount of DNA 28-40 ,ug DNA/g tissue from one year old cane, according to Lin and Walker 1997 was found to be very similar compared by that protocol.

Keywords

• Vitis vinifera L.
• DNA isolation
• RAPD

References

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