Quantitative PCR

Immunohistochemical and molecular analysis of caveolin-1 expression in canine mammary tumors

D. A. P. C. Zuccari, Castro, R., Gavioli, A. F., Mancini, U. M., Frade, C. S., and Leonel, C., Immunohistochemical and molecular analysis of caveolin-1 expression in canine mammary tumors, vol. 11, pp. 153-165, 2012.

Caveolin-1 (Cav-1) is a structural protein present in invaginations of the cell membrane. In human breast cancer, the cav-1 gene is believed to be a tumor suppressor gene associated with inhibition of tumor metastasis. However, little is known about its expression, regulation and function in canine mammary tumors. Expression levels of cav-1 were investigated using real-time PCR and immunohistochemical detection with an anti-human Cav-1 antibody. Gene expression stability of different samples was analyzed using the geNorm software.

Suitable internal control microRNA genes for measuring miRNA abundance in pig milk during different lactation periods

Y. R. Gu, Liang, Y., Gong, J. J., Zeng, K., Li, Z. Q., Lei, Y. F., He, Z. P., and Lv, X. B., Suitable internal control microRNA genes for measuring miRNA abundance in pig milk during different lactation periods, vol. 11. pp. 2506-2512, 2012.

Determination of an optimal set/number of internal control microRNA (miRNA) genes is a critical, but often undervalued, detail of quantitative gene expression analysis. No validated internal genes for miRNA quantitative PCR (q-PCR) in pig milk were available. We compared the expression stability of six porcine internal control miRNA genes in pig milk from different lactation periods (1 h, 3 days, 7 days, 14 days, 21 days, and 28 days postpartum), using an EvaGreen q-PCR approach.

Comparative multiplex dosage analysis in spinocerebellar ataxia type 2 patients

F. Calì, Chiavetta, V., Ragalmuto, A., Vinci, M., Ruggeri, G., Schinocca, P., and Romano, V., Comparative multiplex dosage analysis in spinocerebellar ataxia type 2 patients, vol. 12, pp. 1176-1181, 2013.

We developed a new application of comparative multiplex dosage analysis (CMDA) for evaluation of the ataxin 2 gene. Expansions of the triplet CAG can cause spinocerebellar ataxia type 2 (SCA2), a neurodegenerative disease with an autosomal-dominant mode of inheritance. Molecular diagnosis of SCA2 is routinely based on the use of conventional PCR to detect the CAG expansion. However, PCR does not amplify an allele with an expansion of many triplets (>80), which is typically found in infantile and juvenile forms of SCA2, thus leading to false negatives.

A duplex SYBR Green I real-time quantitative PCR assay for detecting Escherichia coli O157:H7

X. Yang, Cheng, H. W., Chen, L., Zhao, J., Chang, H. T., Wang, X. W., Liu, H. Y., Yao, H. X., Zhang, L. X., and Wang, C. Q., A duplex SYBR Green I real-time quantitative PCR assay for detecting Escherichia coli O157:H7, vol. 12, pp. 4836-4845, 2013.

PCR and hybridization assays are widely used for the detection and identification of Escherichia coli serogroups and serotypes. We used these techniques for the detection of E. coli O157:H7, a dominant serogroup among E. coli strains that are considered major public health problems worldwide. We developed a quantitative PCR assay using SYBR Green I, based on the published sequences of the rfbE and fliC genes from E. coli O157:H7. This method detected the E.

Cloning and quantitative expression analysis of drought-induced genes in soybean

R. Stolf-Moreira, Medri, M. E., Neumaier, N., Lemos, N. G., Brogin, R. L., Marcelino, F. C., de Oliveira, M. C. N., Farias, J. R. B., Abdelnoor, R. V., and Nepomuceno, A. L., Cloning and quantitative expression analysis of drought-induced genes in soybean, vol. 9, pp. 858-867, 2010.

We determined the expression levels of DREB transcription factor (Gmdreb1) and of the genes Gmgols, Gmpip1b, Gmereb, and Gmdefensin in drought-tolerant (MG/BR46-Conquista) and drought-sensitive (BR16) genotypes of soybean, during drought. The trial was carried out in a controlled-environment chamber, set up to provide drought conditions. Sequences of Arabidopsis thaliana DREB-family proteins were used to build a phylogenetic tree through the alignment of the conserved regions near the AP2 domain.

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