Preparing compound heterozygous reference material using gene synthesis technology: a model of thrombophilic mutations

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00179906%3A_____%2F14%3A10283772" target="_blank" >RIV/00179906:_____/14:10283772 - isvavai.cz</a>
Alternative codes found
RIV/00216208:11150/14:10283772
Result on the web
<a href="http://biomed.papers.upol.cz/artkey/bio-201404-0007_preparing_compound_heterozygous_reference_material_using_gene_synthesis_technology_a_model_of_thrombophilic_mu.php#.VK5-h2NWXuc" target="_blank" >http://biomed.papers.upol.cz/artkey/bio-201404-0007_preparing_compound_heterozygous_reference_material_using_gene_synthesis_technology_a_model_of_thrombophilic_mu.php#.VK5-h2NWXuc</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.5507/bp.2014.041" target="_blank" >10.5507/bp.2014.041</a>

Result language
angličtina
Original language name
Preparing compound heterozygous reference material using gene synthesis technology: a model of thrombophilic mutations
Original language description
The aim of our study is to present a novel approach for preparing a compound heterozygous reference material (hetRM) using gene synthesis technology with inverted insertion of wild-type and mutant fragments into a single cloning vector. Factor II (G20210A) and Factor V (G1691A Leiden) gene mutations were used as an experimental model. During the gene synthesis, DNA fragments were aligned in the following order: G1691 FV wild-type forward strain, G20210 FII wild-type forward strain, 1691A FV mutant reverse strain, 20210A FII mutant reverse strain. The complete chain was inserted into a pIDT SMART cloning vector and amplified in an E. coli competent strain. For assessing hetRM characteristics and commutability, we used real-time PCR with subsequent melting curve analysis, real-time PCR with hydrolysis probes, allele-specific amplification, reverse hybridization, and dideoxynucleotide DNA sequencing. All five methods yielded concordant results of DNA analysis of the hetRM. Differences in
Czech name
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Czech description
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Type
J<sub>x</sub> - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)
CEP classification
EB - Genetics and molecular biology
OECD FORD branch
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Project
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Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year
2014
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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