FIGURE 1.
The linker and primer design. (A) The linkers and primers: The 5′ linker is an RNA oligo with 5′OH and 3′OH; the 3′ linker is an adenylylated DNA oligo with 5′App and 3′ ddC (dideoxyC); the RT primer is reverse complementary to the 3′ linker; and the 5′ and 3′ PCR primers partially overlap with the 5′ and 3′ linkers, respectively. (B) The sequence feature of the final cDNA: The amplicon is drawn as a double-stranded DNA with five arrows indicating the primer/linker sequences; the underlined parts are the 13-nt inverted repeats flanking the inserts; and the index represents an 8-nt barcode.
