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High-Resolution Melting Analysis for Identifying Sequence Variations in Nuclear Genes for Assembly Factors and Structural Subunits of Cytochrome C Oxidase

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Part of the Methods in Molecular Biology book series (MIMB, volume 1264)

Abstract

High-resolution melting (HRM) analysis is a simple, sensitive, and cost-effective screening method. HRM enables the detection of homozygous or heterozygous point sequence variants and small deletions within specific PCR products by observing temperature and shape changes in melting curve profiles using fluorescent dyes. Herein, an updated protocol for routine variant screening of nuclear genes encoding assembly factors and structural subunits of cytochrome c oxidase (COX) is described. Nonetheless, the general recommendations given for HRM analysis can be applicable for examining any genetic region of interest.

Key words

COX HRM analysis Screening of sequence variants Melting dye LCGreen Plus LightScanner 
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Notes

Acknowledgements

This work was supported by grants SVV2014/260022, P24/LF1/3 and UNCE 204011 from Charles University in Prague.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of Pediatrics and Adolescent Medicine, First Faculty of MedicineCharles University in Prague and General University Hospital in PraguePrague 2Czech Republic

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