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Cooperative oligonucleotides in purification of cycle sequencing products
KTH, Superseded Departments, Biotechnology.ORCID iD: 0000-0001-8993-048X
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2000 (English)In: BioTechniques, ISSN 0736-6205, E-ISSN 1940-9818, Vol. 29, no 2, 352-+ p.Article in journal (Refereed) Published
Abstract [en]

Nucleic acid hybridization is an essential component in many of today's standard molecular biology techniques. In a recent study, we investigated whether nucleic acid capture could be improved by taking advantage of stacking hybridization, which refers to the stabilizing effect that exists between oligonucleotides when they hybridize in a contiguous tandem fashion. Here, we describe a specific approach for purification of sequencing products using cooperative probes that hybridize to single-strand targets where one of the probes has been coupled to a magnetic bead. This approach has been developed for standard sequencing primers and has been applied to shotgun plasmid libraries. The cooperative probes have been designed to anneal within the common vector sequence and to avoid copurification of non extended sequencing primers and misprimed sequencing products. The reuse of magnetic beads, together with salt independent elution, makes the approach suitable for high-capacity capillary electrophoresis instruments.

Place, publisher, year, edition, pages
2000. Vol. 29, no 2, 352-+ p.
Keyword [en]
gene-expression patterns, c virus-rna, direct capture, dna, microarray
Identifiers
URN: urn:nbn:se:kth:diva-19956ISI: 000088679400031OAI: oai:DiVA.org:kth-19956DiVA: diva2:338648
Note
QC 20100525Available from: 2010-08-10 Created: 2010-08-10 Last updated: 2017-12-12Bibliographically approved

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Uhlén, Mathias

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