Identification of CYP2B6 Sequence Variants by Use of Multiplex PCR with Allele-Specific Genotyping

Identification of CYP2B6 sequence variants by use of multiplex PCR with allele-specific genotyping.

BACKGROUND Cytochrome P450 2B6 (CYP2B6) has a role in the metabolism of many clinically important substances, but the variation within the CYP2B6 gene has not been fully characterized. The aim of the present study was to develop a reliable and robust assay for determining genotypic variants. METHODS We used a two-stage procedure. An initial multiplex PCR reaction amplified the relevant gene f...

Serotype Identification of Recent Iranian Isolates of Infectious Bronchitis Virus by Type-Specific Multiplex RT-PCR

Measles virus genotyping by nucleotide-specific multiplex PCR.

A simple genotyping method based on multiplex PCR has been developed to discriminate between all active measles virus (MV) clades and genotypes (A, B3.1, B3.2, C2, D2-D9, G2-G3, and H1-H2). The sequencing reaction was replaced by six multiplex PCRs: one to identify the clade and five to identify the respective genotype. Primers were sensitive to clade- and genotype-specific nucleotides and gene...

Multiplex allele-specific PCR with optimized locked nucleic acid primers.

The ease and advantages of allelespecific PCR (AS-PCR) for SNP genotyping, along with the difficulty of assay specificity with certain mismatched DNA primers, have been described in prior studies (1–3). More recently, the use of real-time PCR detection formats has been described for one or both alleles of AS-PCR (4–6). These detection enhancement methods do not overcome the inherent difficultie...

Multiplex allele-specific target amplification based on PCR suppression.

We have developed a strategy for multiplex PCR based on PCR suppression. PCR suppression allows DNA target amplification with only one sequence-specific primer per target and a second primer that is common for all targets. Therefore, an n-plex PCR would require only n + 1 primers. We have demonstrated uniform, efficient amplification of targeted sequences in 14-plex PCR. The high specificity of...