title: GSM4835450: HK6_EGF_insensitive; Homo sapiens; miRNA-Seq
library_construction_protocol: Total miRNA was extracted after treatment with Erlotinib using TriFast. 1 ml TriFast was used for each sample to lyse the cells. Cell lysate was mixed with chloroform and centrifuged. Upper aqueous phase was mixed with isopropanol and centrifuged. Supernatant was discarded and pellets were washed with 75% ethanol and centrifuged. Supernatant was again discarded and RNA pellet was resolved in 50 µl RNase-free water. RNA sequencing libraries were generated using the NEBNext Mulitplex Small RNA Library Prep Set for Illumina (NEB E7300S) following size selection by running the samples on a 10 % polyacrylamide gel and isolating the 147 bp. Samples were purified using the QIAquick PCR Purification Kit (Qiagen, 28104). Libraries were validated on a High Sensitivity DNA chip using the 2100 Bioanalyzer (Agilent).