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  • FAQ: Why do I add the SR RT Primer 3 before the 5’ ligation instead of right before the reverse transcription step?

    The SR RT Primer 3 is added before 5’ Ligation because it will hybridize to unligated 3’ SR Adaptor 3 and prevent it from ligating to the 5’ SR Adaptor 3, thus not allowing the creation of adapter-dimer.