title: GSM2065790: MCF7+E2 6h AGO HITS-CLIP (Nested); Homo sapiens; RIP-Seq
library_construction_protocol: Extraction was performed as in Balakrishnan I, et al. Genome-wide analysis of miRNA-mRNA interactions in marrow stromal cells. Stem Cells Dayt Ohio. 2013. Library construction was performed as in Balakrishnan I, et al. Genome-wide analysis of miRNA-mRNA interactions in marrow stromal cells. Stem Cells Dayt Ohio. 2013. with the following modifications: First, a nested reverse transcription primer (5’-CCGCTGGAAGTGACTGA-3’) was used in place of the DP3 primer for all samples. Second, for the "Nested, Protected" samples, phosphorothioate bonds were added between the final four bases of the reverse PCR primer to prevent cleavage by exonuclease activity of the polymerase (5’-GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCCCGCTGGAAGTGACTGA*C*A*C-3’, “*” indicate phosphorothioate bonds). The second PCR was performed with index primers compatible with the new first-stage primers (5’-CAAGCAGAAGACGGCATACGAGATCGGT NNNNNN GTGACTGGAGTTCAGACGTGTGCTCTTCCGATC-3’).
attributes.tag: GEO Accession
attributes.value: GSM2065790