Similar results were observed with our synthetic TTV miRNA candidates and corresponding reporter constructs (Figure S1C). Therefore, we conclude that the TTV miRNAs are active in RISC. Taken together, we have identified the general pathway through which biologically active miRNAs are generated. Due to smaller size of sensitive element, the SPRi is usually in some degree less sensitive than the standard SPR; however, the use of enhancement (like nanoparticles) makes it possible to detect nucleotide fragments down to femtomolar concentrations. Total RNA was harvested with PIG-B at thirty hours posttransfection. RNA was treated with DNase (Qiagen) and purified using RNeasy Minielute kit (Qiagen). RNA integrity was verified on a 1.0% agarose (Formaldehyde, MOPS) denaturing gel. Sequences were mapped to the synthetic region of the AFL351132 NCBI reference sequence.
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