Rna and mirna extraction kits

Brandon University Research Ethics Committee approves this study (#21986, 21 April 2017). The human tissues used in this project were obtained by Dr. Peeyush K Lala at the University of Western Ontario (UWO) from the Ontario Institute for Cancer Research (OICR) repository (created on the basis of donor consent) following approval of human ethics by the Ethics Review Board of the OICR and UWO. Total RNA and miRNA were extracted using Qiagen (Qiagen, Toronto, ON, Canada) RNA and miRNA extraction kits followed by cDNA synthesis at UWO, and a portion of cDNA of all samples were transferred to Dr. Mousumi Majumder at Brandon University following a material transfer agreement.

To examine the clinical relevance of miR655 expression in breast cancer, we obtained frozen female human breast tumor (n = 105) and control (n = 20) tissues (adjacent non-tumor tissue from unrelated patients) from the Ontario Tumor Bank, maintained by the Institute for Cancer Research (OICR). Demographic description of the sample set is described earlier^{23 ,37} . We used Qiagen RNA and miRNA extraction kits to extract mRNA/miRNA from tissue followed by cDNA synthesis and Taqman miRNA and gene expression assays (miR655, COX-2, EP4 and ALDH1A) as described before^{23 ,37} . Negative ∆Ct values (≤0) are indicative of higher miRNA expression in tissue samples.