GSM2598375: SCST2; Homo sapiens; miRNA-Seq
mirVana miRNA isolation kit (Applied Biosystems/ Ambion, Austin, TX) was used to extract small RNA for cloning small RNAs were ligated with adenylated 3’-adaptor oligonucleotide [/5rApp/CTGTAGGCACCATCAAT/3ddC; IDT Inc., Coralville, IA, USA] and purified on a 12% denaturing polyacrylamide gel. Second ligation reaction was performed with 5’-adaptor oligonucleotide [5'-ACGCTCTTCCGATCTv-3' (uppercase, DNA; v = barcodes with triple RNA molecules: aaa, ggg, ccc or uuu; IDT)], followed by gel purification. The double ligated product was converted to cDNA using the reverse transcription enzyme SuperScript II (Invitrogen) and the reverse primer [5'-ATTGATGGTGCCTACAG-3'], and then PCR amplified for 16-20 cycles using a forward primer 5'-GAT ACG GCG ACC ACC GAG ATC TAC ACT CTT TCC CTA CAC GAC GCT CTT CCG ATC T-3' and a reverse primer 5'-CAA GCA GAA GAC GGC ATA CGA GCT CTT CCG ATC TAT TGA TGG TGC CTA CAG- 3'.
GEO Accession
GSM2598375