http://www.cnr.it/ontology/cnr/individuo/prodotto/ID26767
Hexaprimer amplification refractory mutation system PCR for simultaneous single-tube genotyping of 2 close polymorphisms (Articolo in rivista)
- Type
- Label
- Hexaprimer amplification refractory mutation system PCR for simultaneous single-tube genotyping of 2 close polymorphisms (Articolo in rivista) (literal)
- Anno
- 2008-01-01T00:00:00+01:00 (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi
- 10.1373/clinchem.2007.095703 (literal)
- Alternative label
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
- Piccioli P; Serra M; Pedemonte S; Balbi G; Loiacono F; Lastraioli S; Gargiulo L; Morabito A; Zuccaro D; Del Mastro L; Pistillo MP; Venturini M; De Angioletti M; Notaro R (literal)
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- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo
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- ISI Web of Science (WOS) (literal)
- Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
- Laboratory of Human Genetics Medical Oncology C, Laboratory of Translational Research A,
Medical Oncology A IST, Istituto Nazionale per la Ricerca sul Cancro Genoa, Italy;
Department of Oncology, Biology and Genetics University of Genoa, Genoa, Italy;
Medical Oncology Ospedale Sacro Cuore - Don Calabria Verona, Italy;
Istituto di Genetica e Biofisica \"Adriano Buzzati Traverso\" (IGB-CNR) Naples, Italy;
Laboratory of Genetics and Gene Transfer Core Research Laboratory - Istituto Toscano Tumori (CRL-ITT)
Florence, Italy (literal)
- Titolo
- Hexaprimer amplification refractory mutation system PCR for simultaneous single-tube genotyping of 2 close polymorphisms (literal)
- Abstract
- Tetraprimer amplification refractory mutation system PCR (T-ARMS-PCR) is a simple and inexpensive genotyping method for differentiating both alleles of a polymorphism/mutation (both single-nucleotide polymorphisms and small insertions/deletions) with a single-tube PCR (1). In T-ARMS-PCR, a pair of common (outer) primers produces a non-allele-specific control amplicon and in combination with 2 allele-specific (inner) primers (designed to anneal in the opposite orientation) produces 2 allele-specific amplicons. These allele-specific amplicons have different sizes because the polymorphism/mutation is asymmetrically located with respect to the common primers. Thus, the amplicons can be separated by standard gel electrophoresis. T-ARMS-PCR has also been designed in a multiplex fashion to genotype more than one polymorphism/mutation by a single-tube PCR (2).
We describe a modified multiplex T-ARMS-PCR, the hexaprimer ARMS-PCR (H-ARMS-PCR), which is for when 2 polymorphisms are close in the sequence. H-ARMS-PCR uses only 6 primers and provides direct information about haplotype structure.
The CTLA4 gene (cytotoxic T-lymphocyte-associated protein 4; also known as CD152) is a negative regulator of T-cell function (3). The CTLA4 polymorphisms -318 C>T (rs5742909) and +49 A>G (rs231775) are associated with susceptibility to autoimmune diseases and cancer (3)(4). To genotype these 2 polymorphisms, which are only 365 bp apart in the 5? region of the CTLA4 gene, we designed an H-ARMS-PCR that combines a single pair of common primers and 2 pairs of allele-specific primers in the same tube (Fig. 1A? ). (literal)
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