Short template amplicon and multiplex megaprimer-enabled relay (STAMMER) sequencing, a simultaneous approach to higher throughput sequence-based typing of polymorphic genes View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2010-04

AUTHORS

Chrissy h. Roberts, Neema P. Mayor, J. Alejandro Madrigal, Steven G. E. Marsh

ABSTRACT

Sequence-based typing (SBT) is a powerful method of genotyping in highly polymorphic gene systems. In standard SBT methods, both strands of a double-stranded template amplicon are sequenced in separate reactions in order to achieve high quality data across the region of interest. The amount of informative data that is obtained from the second strand sequence is often low, whilst the impact of performing second strand sequencing on costs and throughput are significant. Here we present short template amplicon and multiplex megaprimer-enabled relay (STAMMER) sequencing, a novel simultaneous sequence-based typing methodology that allows the detection of any practical amount of useful sequence from a plurality of distinct polymerase chain reaction products in a single sequencing reaction. In addition to simultaneous bidirectional sequencing, we show how the STAMMER approach can be used to simultaneously sequence a number of regions of interest that are not physically linked within the range of a single sequencing reaction. The efficiencies of this method could impact significantly on the output of SBT laboratories. More... »

PAGES

253-260

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00251-010-0432-5

DOI

http://dx.doi.org/10.1007/s00251-010-0432-5

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1015780824

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/20204613


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