GSM5350786: RF_GC_rep3; Gallus gallus; RNA-Seq
5-6 recently selected follicles (6-8 mm) per bird were removed and granulosa cells were collected and pooled. Total RNA was extracted from granulosa cells using a RNeasy Mini kit with optional on-column DNase treatment (Qiagen Inc., Valencia CA). Quantity and purity of the samples was analyzed using spectrophotometry. RNA integrity was then determined by Fragment Analyzer (Advanced Analytical, Ames IA). All samples had a RQN of >9.9. Samples were enriched by PolyA+ RNA isolation using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs, Ipswich MA). Libraries were then generated using the NEBNext Ultra II [Directional] RNA Library Prep Kit (New England Biolabs, Ipswich MA).