title: GSM2425636: MDA231-UCA1-MS2-RIP; Homo sapiens; miRNA-Seq
library_construction_protocol: Cells were lysed in an ice-cold lysis buffer (20mM Tris•Cl pH7.5, 50mM NaCl, 5 mM MgCl2, 0.5% NP-40) containing protease inhibitors and RNasin (200 U/mL).Lysates were centrifuged at 12,000 rpm at 4°C for 15 minutes, to remove cell debris.Supernatants were incubated with Amylose-resin-attached MBP-MCP (MS2 coat protein) at 4°C for 5 h with gentle rotation. After incubation, the supernatant was removed by a short centrifugation and the Amylose beads were washed three times in lysis buffer，Bound UCA1-MS2 RNP complexes were eluted with 100 µl lysis buffer containing 20 mM maltose. miRNAs associated with UCA1 were extracted and analyzed by miRNA-seq. Libaries were prepared for sequencing according to Illumina's protocol. The experiment was performed by the Guangzhou RiboBio co. Ltd in Guangzhou, China.
attributes.tag: GEO Accession
attributes.value: GSM2425636