Figure 4.

Scheme of Argonaute high-throughput sequencing of RNAs by cross linking and immunoprecipitation. RNA and protein are cross linked by UV and then RNA is digested by RNase, a treatment which is finally immunoprecipated. 5′ ends are dephosphorylated and 3′ ends are adapter ligated followed by phosphate addition at 5′ ends. The complexes of RNA and protein are separated by SDS-PAGE and RNA are amplified after 5′-adaptor ligation. These amplified products will be sequenced by next generation sequencers and finally computational approaches will help identify the miRNA target.