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New universal primers facilitate PyrosequencingTM

Item Type:Article
Title:New universal primers facilitate PyrosequencingTM
Creators Name:Aydin, A. and Toliat, M.R. and Baehring, S. and Becker, C. and Nuernberg, P.
Abstract:A cost-driving factor in Pyrosequencing is the need for single-stranded PCR products that are usually obtained by biotin-labeling of one primer. We designed new universal primers that allow the introduction of biotin during the specific PCR at either the forward or the reverse primer in a single reaction. When converting five human single nucleotide polymorphism assays from the standard format into the universal format, we obtained pyrograms of similar good quality. Although the universal nonhuman sequences are unlikely to form loop structures, we mostly failed to establish assays without using the proprietary software from Biotage AB. Nevertheless, our universal primer pair adds flexibility to the process of assay design due to optional strand selection and contributes to the reduction of genotyping costs.
Keywords:Optional Strand Selection, Pyrosequencing, Universal Primers
Source:Electrophoresis
ISSN:0173-0835
Publisher:Wiley (Germany)
Volume:27
Number:2
Page Range:394-397
Date:January 2006
Official Publication:https://doi.org/10.1002/elps.200500467
PubMed:View item in PubMed

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