Design Description | At first, 5'/3' adaptors were ligated to the both ends of shortRNAs.
The structure is
"5'-ACGCTCACAGAATTCAAA-shortRNA(16-51nt)-U-barcode(4nt)-GAATTGTCACGAGGCCAGCGTGC-3'".
After RT reaction and PCR amplification, the PCR products were restricted by EcoRI
to make cohesive ends on the both ends of shortRNA tags. The structure is
"5'-AATTCAAA-shortRNA(16-51nt)-T-barcode(4nt)-G-3'". These shortRNA tags were
concatenated to make long fragments, and then the 454 adaptors were ligated to the
both ends of concatemers. The number and the direction of concatenated shortRNA tags
are various. |