Small RNA Sequencing of non-coding RNAs which are less than 200 nucleotides extracted from LNCaP, human prostate cancer cell and HCT116, human colorectal cancer

This project investigated the specific interactions between non-coding RNAs extracted from cell and compounds. Using McCoy's 5A medium (FBS 10%), we subcultured LNCaP, a human prostate cancer cell, four times and HCT116, a human colorectal cancer cell, two times. For each Petri dish, cells were dissolved in solution D which contains guanidinium for ribonuclease inhibition, and preserved at -80°C. After total RNA extraction according to acid-guanidinium-phenol-chloroform method, small RNAs with less than 200 nucleotides were purified through columns. By binding assay with NHS beads attached compound, compound binding to small RNAs from LNCaP were extracted. We prepared 5 LNCaP RNA samples: compound binding RNA samples for each 3 compounds, native LNCaP small RNA sample, and beads binding RNA sample. In addition, we prepared native HCT116 small RNA sample. Sequence libraries were prepared using Illumina's TruSeq small RNA Library Kit SetA, and the sequencing was performed with Illumina's MiSeq.