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Reference Barcoded primers used in multiplex amplicon pyrosequencing bias amplification. Berry D, Ben Mahfoudh K, Wagner M, Loy A. Applied and environmental microbiology. 2011.
Abstract "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 pyrosequencing libraries.
Pubmed ID 21890669
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