Is total RNA extraction possible from small volume goat milk (7.5 mL) for RT-qPCR studies? RT-qPCR çalışmaları için küçük hacimli keçi sütlerinden (7.5 mL) total RNA ekstraksiyonu mümkün müdür?

ERASLAN ŞAKAR A., ÖZKAN H., YAKAN A.

Ankara Universitesi Veteriner Fakultesi Dergisi, cilt.65, sa.1, ss.57-61, 2018 (SCI-Expanded) identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 65 Sayı: 1
  • Basım Tarihi: 2018
  • Doi Numarası: 10.1501/vetfak_0000002827
  • Dergi Adı: Ankara Universitesi Veteriner Fakultesi Dergisi
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.57-61
  • Anahtar Kelimeler: Goat, Milk volume, RNA extraction, RT-qPCR
  • Hatay Mustafa Kemal Üniversitesi Adresli: Evet

Özet

This study aimed to study the application availability of RT-qPCR by total RNA extraction from small volumes of milk samples. RNA was extracted from 7,5 mL milk. Amount and quality of RNA and Ct results obtained via RT-qPCR were compared with the values obtained from 100 mL milk. A260/A280 ratio of absorbance was found to be higher than 1,70 in all samples. Mean RNA purity were identified as 1,87±0,07 and 1,77±0,01 in 7,5 mL and 100 mL milk samples respectively. While mean total RNA concentration was 149,37±43,40 μg/mL in 7,5 mL milk sample, it was measured to be 309,03±77,82 μg/mL in 100 mL milk sample. Mean somatic cell count (SCC) of the 8 goats used in the study was calculated as 486,37±185,68 x 103 cells/mL. Ribosomal Protein Lateral Stalk Subunit P0 (RPLP0) gene which is accepted as the reference gene in goat somatic cells was used to test application availability of obtained RNA in RT-qPCR. Mean Ct results were found to be 20,33±0,39 and 19,51±0,41 in 7,5 mL and 100 mL groups respectively, creating no difference between the two groups (P = 0,170). Quality and amount of obtained RNA and Ct results showed that it was possible to extract RNA by using 7,5 mL milk.