Barcoded primers used in multiplex amplicon 5 pyrosequencing bias amplification
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Barcoded primers used in multiplex amplicon pyrosequencing bias amplification.
"Barcode-tagged" PCR primers used for multiplex amplicon sequencing generate a thus-far-overlooked amplification bias that produces variable terminal restriction fragment length polymorphism (T-RFLP) and pyrosequencing data from the same environmental DNA template. We propose a simple two-step PCR approach that increases reproducibility and consistently recovers higher genetic diversity in pyro...
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