PCR strategy and primers used to clone PMP-1a

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  • a The PCR primer pairs were used as indicated by the numbered arrows: 3′-RACE, primers 1 and 2; 5′-RACE, primers 3 to 6; full-length PCR, primers 7 and 8. Note that primer 3 contains deoxyinosine and a degeneracy of 32-fold. 5′-RACE adaptor primers are specific for an adaptor sequence ligated to decapped full-length mRNA sequences. 5′-RACE was conducted with outer and inner (nested) adaptor primers paired with gene-specific primers (GSPs). Shading is as follows: hatched, 5′ adaptor; light gray, untranslated regions; dark gray, signal peptide domain; black, mature peptide domain; AAAAA, poly(A) tail.