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High‐throughput mutational screening for beta‐thalassemia by single‐nucleotide extension
Author(s) -
Galbiati Silvia,
Chiari Marcella,
Macellari Micol,
Ferrari Maurizio,
Cremonesi Laura,
Cretich Marina
Publication year - 2007
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200700181
Subject(s) - genotyping , throughput , dna sequencer , beta thalassemia , high throughput screening , mutation , biology , computational biology , beta (programming language) , thalassemia , genetics , dna , microbiology and biotechnology , gene , dna sequencing , genotype , computer science , telecommunications , wireless , programming language
Abstract In this work a high‐throughput method based on the single‐nucleotide extension (SNE) reaction and multicolour detection in a DNA sequencer was developed to screen for eight mutations in the human beta‐globin gene: IVSI.110, cd39, IVSI.1, IVSI.6, IVSII.745, HbC, HbS and cd6. The method has been validated on a large number of samples for the two most common mutations causing beta‐thalassemia in the Mediterranean area (IVSI.110 and cd39). The development of a high‐throughput, fast and reliable method to assay beta‐thalassemia mutations represents a significant improvement in molecular diagnosis of this disease. The multicolour detection and the use of multiple injections further enhances the throughput of mutational screening by the DNA sequencer and facilitates automated genotyping for routine molecular diagnostics.