GSM9056006: Mex3a_cKO-Late_Polysome-RNA_rep2; Mus musculus; RNA-Seq
Tissue lysates were loaded on top of a sucrose gradient and subjected to ultracentrifugation. Approximately 42 350uL fractions were collected along the sucrose gradient where trop fractions were less dense and bottom fractions were more dense. Early polysomes were from fractions 27-34 and Late polysomes were from fractions 35-42. 50uL from each fraction was pooled and RNA was extracted using Trizol LS reagent and 1-Bromo-3-chloropropane. After ETOH was added to samples, the Zymo RNA Clean and Concentrator kit was used to finalize extraction protocol. libraries were prepared with SMARTer Stranded Total RNA-Seq Kit v2 - Pico Input Mammalian kit (Takara)