GSM8321165: D5; synthetic construct; SELEX
RAPID-SELEX using PARP1 and MeCP2 and an RNA library (independent experiments for each protein) were used to select for RNAs that are high binders for this proteins. Proteins were bound to Ni-NTA-micro-columns and incubated with RNA library for 12 rounds. Each round consist of a non-amplification and an amplification cycles (after reverse transcribing of RNA to cDNA) to generate the selected RNAs with enriched specificity for next round of selection. The RNA library was containing ~ 5 × 10^15 unique 120-mer sequences consisting of a random 70-mer flanked by constant regions at each end [5' AAGCTTCGTCAAGTCTGCAGTGAA-N70-GAATTCGTAGATGTGGATCCATTCCC]. The RNA library contained (on average) of 4 to 6 copies of each unique sequence. RNAs from RAPID-SELEX rounds 3, 8, and 12, were cDNA converted and PCR amplified to form dsDNAs. These DNAs were subjected to to Illumina Hi-Seq 4000 platform according to the manufacturer's instructions.