PCR

Expression and significance of miR-21 in multiple myeloma patients

J. H. Wang, Zhou, W. W., Liu, B. X., Man, D. L., Yang, Z. D., Liu, F. R., Shang, H., Wang, J. H., Zhou, W. W., Liu, B. X., Man, D. L., Yang, Z. D., Liu, F. R., Shang, H., Wang, J. H., Zhou, W. W., Liu, B. X., Man, D. L., Yang, Z. D., Liu, F. R., and Shang, H., Expression and significance of miR-21 in multiple myeloma patients, vol. 15, p. -, 2016.

The aim of the present study is to examine the expression level of peripheral mir-21 in multiple myeloma (MM) patients and to determine its clinical significance. MM patients (30), monoclonal gammopathy of undetermined significance (MGUS) patients (14), and normal controls (20) were recruited to determine the serum level of β2-MG, IgA and IgM, IgG, λ, κ, TP, ALB, Hb, LDH, and Ca2+. Gene expression of mir-21 was quantified by SYBR green real-time fluorescent quantitative PCR.

Restriction-ligation-free (RLF) cloning: a high-throughput cloning method by in vivo homologous recombination of PCR products

Y. Wang, Liu, Y., Chen, J., Tang, M. J., Zhang, S. L., Wei, L. N., Li, C. H., and Wei, D. B., Restriction-ligation-free (RLF) cloning: a high-throughput cloning method by in vivo homologous recombination of PCR products, vol. 14, pp. 12306-12315, 2015.

In this study, we optimized a restriction-ligation-free (RLF) method to save time and cost of constructing multiple plasmids with the same gene insert, and examined the efficacy of RLF on high-throughput multi-plasmid cloning. This method utilizes the precise DNA repair and recombination systems within Escherichia coli, which allows to bypass the in vitro restriction and ligation enzyme reactions commonly included in routine cloning procedures. A homologous arm is linked to the 5'-end of the forward primer used to amplify both the target gene and vector.

Molecular analysis of patients suspected of Fragile X Syndrome

A. P. Amancio, Melo, C. Ade O., A. Vieira, deM., Minasi, L. B., D. Silva, deM. e, da Silva, C. C., and da Cruz, A. D., Molecular analysis of patients suspected of Fragile X Syndrome, vol. 14, pp. 14660-14669, 2015.

The aim of this study was to validate the molecular genetic diagnosis of patients suspected of Fragile X Syndrome (FXS) in the Laboratory of Human Cytogenetics and Molecular Genetics (LaGene) of the Department of Health of the State of Goiás, using polymerase chain reaction (PCR). Thirty-five patients referred by public health doctors to LaGene, indicating clinical diagnosis of FXS, were selected for this study. Two PCR analyses were performed using different primers, one for screening (PCR-T) and one for the detection of the pre-mutation (PCR-P).

Genetic relatedness of artichoke (Cynara scolymus L.) hybrids using random amplified polymorphic DNA (RAPD) fingerprinting

M. A. Sharaf-Eldin, Al-Tamimi, A., Alam, P., Elkholy, S. F., and Jordan, J. R., Genetic relatedness of artichoke (Cynara scolymus L.) hybrids using random amplified polymorphic DNA (RAPD) fingerprinting, vol. 14, pp. 18431-18439, 2015.

The artichoke (Cynara scolymus L.) is an important food and medicinal crop that is cultivated in Mediterranean countries. Morphological characteristics, such as head shape and diameter, leaf shape, and bract shape, are mainly affected by environmental conditions. A molecular marker approach was used to analyze the degree of polymorphism between artichoke hybrid lines. The degree of genetic difference among three artichoke hybrids was evaluated using random amplified polymorphic DNA-PCR (RAPD-PCR).

Study of the genetic diversity and structure of a natural population of Nectandra megapotamica (Spreng.) Mez. using RAPD markers

L. S. Costa, Reiniger, L. R. S., Heinzmann, B. M., Amaral, L. P., and Serrote, C. M. L., Study of the genetic diversity and structure of a natural population of Nectandra megapotamica (Spreng.) Mez. using RAPD markers, vol. 14, pp. 18407-18413, 2015.

Nectandra megapotamica (Spreng.) Mez. is a tree species that naturally occurs in the Atlantic Forest, Brazil. The aim of this study was to evaluate the genetic diversity and structure of a natural population of 12 N. megapotamica individuals using random amplified polymorphic DNA markers. Eleven primers were used in this study, producing 81 bands, of which 98.99% were polymorphic. Analysis using STRUCTURE defined three different clusters (K = 3), results that were consistent with those of principal coordinates analysis.

Detection of Toxoplasma gondii DNA in naturally infected sheep’s milk

deSantana D. Rocha, Moura, R. Lde Sousa, Maciel, B. M., Guimarães, L. A., O’dwyer, H. N. S., Munhoz, A. D., and Albuquerque, G. R., Detection of Toxoplasma gondii DNA in naturally infected sheep’s milk, vol. 14, pp. 8658-8662, 2015.

The objective of this study was to verify whether Toxoplasma gondii is excreted in the milk of naturally infected sheep. In order to accomplish this, 275 lactating ewes were used; these were bred extensively in 17 estates distributed across nine cities. Polymerase chain reaction amplification was used to detect T. gondii DNA in milk samples, and the indirect immunofluorescence test was employed for the detection of anti-T. gondii IgG antibodies in the sera, with a cut-off value of 1:64.

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