Figure 1
From: Parallel DNA pyrosequencing unveils new zebrafish microRNAs
Outline of the experimental protocol used for preparation of small RNA libraries. RNA was isolated from ZF developmental samples and from adult tissues using TRIzol® and fractionated on 12.5% denaturing PAGE. Small RNAs were purified from those gels and then ligated to a 3' adapter (AMP-5'p-5'p/CTGTAGGCACCATCAATdi-deoxyC- 3') and to a 5' linker (see Methods). cDNA was prepared and amplified using 20 PCR cycles. PCR products were subjected to clonal amplification by emulsion PCR and then pyrosequenced using a 454 genome sequencer.