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. 1994 Mar 15;91(6):2245–2249. doi: 10.1073/pnas.91.6.2245

Manifold sequencing: efficient processing of large sets of sequencing reactions.

A Lagerkvist 1, J Stewart 1, M Lagerström-Fermér 1, U Landegren 1
PMCID: PMC43347  PMID: 8134382

Abstract

Automated instruments for DNA sequencing greatly simplify data collection in the Sanger sequencing procedure. By contrast, the so-called front-end problems of preparing sequencing templates, performing sequencing reactions, and loading these on the instruments remain major obstacles to extensive sequencing projects. We describe here the use of a manifold support to prepare and perform sequencing reactions on large sets of templates in parallel, as well as to load the reaction products on a sequencing instrument. In this manner, all reaction steps are performed without pipetting the samples. The strategy is applied to sequencing PCR-amplified clones of the human mitochondrial D-loop and for detection of heterozygous positions in the human major histocompatibility complex class II gene HLA-DQB, amplified from genomic DNA samples. This technique will promote sequencing in a clinical context and could form the basis of more efficient genomic sequencing strategies.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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