Barcoded Primers Used in Multiplex Amplicon Pyrosequencing Bias Amplification

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...

Barcoded primers used in multiplex amplicon 5 pyrosequencing bias amplification

Comparative Analysis of Korean Human Gut Microbiota by Barcoded Pyrosequencing

Human gut microbiota plays important roles in harvesting energy from the diet, stimulating the proliferation of the intestinal epithelium, developing the immune system, and regulating fat storage in the host. Characterization of gut microbiota, however, has been limited to western people and is not sufficiently extensive to fully describe microbial communities. In this study, we investigated th...

Analysis of Microbial Diversity by Amplicon Pyrosequencing

Selecting Degenerate Multiplex PCR Primers

Single Nucleotide Polymorphism (SNP) Genotyping is an important molecular genetics technique in the early stages of producing results that will be useful in the medical field. One of the proposed methods for performing SNP Genotyping requires amplifying regions of DNA surrounding a large number of SNP loci. In order to automate a portion of this method and make the use of SNP Genotyping more wi...