GSM4041420: RPs7_Gard_2; Macaca mulatta; RNA-Seq
We separated the serum and cellular fractions, lysed and discarded the red cells from the cell pellet with red blood cell lysis buffer (RBC lysis solution, 5 Prime Inc.), and lysed the remaining white blood cell fraction in Qiazol for storage at -80°C. We extracted total RNA from each sample using the Qiagen miRNAEASY kit. Each RNA-sequencing library was prepared from 200 ng of total RNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module and the NEBNext Ultra RNA Library Prep Kit (New England Biolabs), following the manufacturer's instructions and selecting for ~350 bp size fragments. Libraries were amplified via PCR for 13 cycles, barcoded, and pooled into sets of 10-12 samples for sequencing on an Illumina HiSeq 2500.