GSM4838130: CF ALI Polyps cells - CF donor PCF4; Homo sapiens; miRNA-Seq
Total RNA were extracted using TRIzol according to manufacturer's instructions Libraries were prepared according to Illumina's instructions accompanying the TruSeq Small RNA kit (part# RS-200-0012). One μg of total RNA with a RIN>7 was used for library construction. After ligation of adapters to the 3'- and 5'-ends, RNA were reverse transcribed using Superscript II and amplified with barcoded primers. Correct enrichment of cDNA containing miRNA sequences was controlled by running each sample on a High Sensitivity DNA Chip on a Bioanalyzer 2100. cDNA derived from microRNAs were selected by purification through a 6% TBE-polyacrylamide gel. After validation of the size using a Bioanalyzer 2100, 12 small-RNA libraries were pooled together at equal molarity to create a unique pool and sequenced on a MiSeq