GSM5641666: 53C; Homo sapiens; RNA-Seq
Samples were collected in 250 uL of Total RNA lysis buffer and RNA was extracted according to manufacturer's instructions from the GenElute™ Mammalian Total RNA Miniprep Kit (Sigma, RTN350-1KT). RNA was quantified on a Nanodrop machine. Libraries were prepared with the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina using the NEBNext Poly(A) mRNA Magnetic Isolation Module (NEB #E7490). Briefly, 300 ng of RNA were used to start with; these were incubated for 10 minutes at 94C to fragment the RNA as in step 1.2.37. 10 cycles were used for the indexing PCR (step 1.9.3). Other than the above, all steps were conducted as per the protocol.