Validação de primers para análises quantitativas por PCR em tempo real dos genes FSHR, LHR e HAS2 em tecidos caprinos / Primers validation for quantitative real-time PCR analysis of FSHR, LHR and HAS2 genes in caprine tissues
Freitas, Jeferson Lucas Sousa; Romcy, Kalil Andrade Mubarac; Viana, Ana Clara Negreiros Parente Capela Sampaio; Freitas, Vicente José de Figueirêdo; Teixeira, Dárcio Ítalo Alves; Melo, Luciana Magalhães.
R. bras. Reprod. Anim.;
40(4): 489-490, Out-Dez. 2016.
Artigo
em Português
| VETINDEX
| ID: vti-24243
Resumo
This study aimed to validate primers to amplify FSHR, LHR and HAS2 genes for quantitative analysisin goat. Thus, total RNA samples were extracted from both goat and bovine cumulus cells and reversetranscripted to obtain cDNA. Specific primer pairs were designed using Primer-BLAST software. First, primerconcentrations were adjusted to achieve the lowest Ct (threshold cycle). Subsequently, standard curves wereconstructed by serial dilution of cDNA. After primers concentrations adjustment, all primer pairs producedspecific amplicons in the presence of the target DNA. The optimized concentration of primer pairs was 1.0 μMand 0.8 μM for primers HAS2-1 and HAS2-2, and 3.0 μM for FSHR and LHR pair of primers. The primerefficiencies accessed by standard curves were: 1.01 for HAS2-1; 1.15 for HAS2-2; 1.02 for FSHR and 0.98 forLHR. In conclusion, all primers designed were appropriate for quantitative gene expression analysis purposes.(AU)
Biblioteca responsável:
BR68.1
Localização: BR68.1
Texto completo