PCR

Detection of genetically modified maize and soybean in feed samples

S. Meriç, Çakır, Ö., Turgut-Kara, N., and Arı, Ş., “Detection of genetically modified maize and soybean in feed samples”, vol. 13, pp. 1160-1168, 2014.

Despite the controversy about genetically modified (GM) plants, they are still incrementally cultivated. In recent years, many food and feed products produced by genetic engineering technology have appeared on store shelves. Controlling the production and legal presentation of GM crops are very important for the environment and human health, especially in terms of long-term consumption. In this study, 11 kinds of feed obtained from different regions of Turkey were used for genetic analysis based on foreign gene determination.

Comparison of methods for high quantity and quality genomic DNA extraction from raw cow milk

T. Usman, Yu, Y., Liu, C., Fan, Z., and Wang, Y., “Comparison of methods for high quantity and quality genomic DNA extraction from raw cow milk”, vol. 13, pp. 3319-3328, 2014.

Isolation of sufficient quantities of high quality DNA is a prerequisite for molecular studies. Milk somatic cells can be used; however, inhibitors such as fats and proteins make milk a difficult medium for extracting large amounts of quality DNA. We optimized, evaluated and compared three methods, Modified Nucleospin Blood Kit method, Modified TianGen Kit method and Phenol-Chloroform method for genomic DNA extraction from bovine milk. Individual cows’ milk and bulk milk samples were collected from a China agricultural university dairy farm.

Comparative study of DNA extraction methodologies from goat sperm and its effects on polymerase chain reaction analysis

E. C. B. Silva, Pelinca, M. A., Acosta, A. C., Silva, D. M. F., Filho, M. A. Gomes, and Guerra, M. M. P., “Comparative study of DNA extraction methodologies from goat sperm and its effects on polymerase chain reaction analysis”, vol. 13, pp. 6070-6078, 2014.

Successful DNA extraction is indispensable for molecular methods based on polymerase chain reaction (PCR); however, goat sperm DNA extraction is limited. Thus, the aim of this study was to evaluate three methods to extract DNA from goat sperm for use in PCR. Eight goat semen pools were used for DNA extraction by using the DNeasy Blood & Tissue Kit, phenol-chloroform, and Chelex-100 methods.

Single- and double-SSR primer combined analyses in rice

J. Ma, Guan, S. C., Zhang, Z., and Wang, P. W., “Single- and double-SSR primer combined analyses in rice”, vol. 11, pp. 1032-1038, 2012.

Polymerase chain reaction (PCR) is the foundation of SSR molecular marker technology. We used sib rice varieties J518, XD1 and SD23 as experimental materials, selecting 30 pairs of SSR primers, including RM127, RM337 and RM5172, covering the rice genome, and performed single- and double-SSR primer combined analyses. We found that under the same PCR system and conditions, a single primer of the SSR primer pairs could amplify the same fragments as double primers do.

Detection limits of the strip test and PCR for genetically modified corn in Brazil

V. E. Nascimento, Von Pinho, E. V. R., Von Pinho, R. G., and Júnior, A. Ddo Nascime, “Detection limits of the strip test and PCR for genetically modified corn in Brazil”, vol. 11, pp. 2497-2505, 2012.

Brazilian legislation establishes a labeling limit for products that contain more than 1% material from genetically modified organisms (GMOs). We assessed the sensitivity of the lateral flow strip test in detection of the GMO corn varieties Bt11 and MON810 and the specificity and sensitivity of PCR techniques for their detection. For the strip test, the GMO seeds were mixed with conventional seeds at levels of 0.2, 0.4 and 0.8% for Bt11, and 0.4, 0.8 and 1.6% for MON810. Three different methodologies were assessed and whole seeds, their endosperm and embryonic axis were used.

A novel missense mutation in exon 7 of the ECM1 gene in an Iranian lipoid proteinosis patient

F. Izadi, Mahjoubi, F., Farhadi, M., Tavakoli, M. M., and Samanian, S., “A novel missense mutation in exon 7 of the ECM1 gene in an Iranian lipoid proteinosis patient”, vol. 11, pp. 3955-3960, 2012.

Lipoid proteinosis (LP) is a rare autosomal recessive disorder. Classical clinical features include warty skin infiltration, papules on the eyelids, skin scarring, as well as extracutaneous abnormalities such as hoarseness of the voice, epilepsy, and neuropsychiatric abnormalities. A defect in the ECM1 gene is responsible for this disease. A 21-year-old female patient from consanguineous parents (first cousins) was referred to our clinic with many symptoms of LP, such as hoarse voice from infancy, diffuse acneiform scars on her face, and hyperkeratosis on her knees and elbows.

Polymorphisms in the ovine myostatin gene are associated with birth weight but not with weight gain in Iranian Makoei sheep

M. Farhadian, Hashemi, A., Mardani, K., Darvishzadeh, R., and Jafari, S., “Polymorphisms in the ovine myostatin gene are associated with birth weight but not with weight gain in Iranian Makoei sheep”, vol. 11, pp. 3568-3575, 2012.

Myostatin, a transforming growth factor-beta superfamily member, has been well documented as a negative regulator of muscle growth and development. Myostatin, which has 376 amino acids, is synthesized as a precursor protein. Polymorphism of the myostatin gene in Makoei sheep was investigated by PCR and single-strand conformation polymorphism technique (SSCP). Genomic DNA of 92 sheep was isolated from whole blood. A 417-bp myostatin intron I segment was amplified by standard PCR, using locus-specific primers.

Genetic variability in the ITS and IGS regions of the ribosomal DNA of Acremonium cavaraeanum exhibiting antimicrobial activity

A. R. Todaro, Nascimento, V. X., Souza, N. C. C., Silva, P. P., Santos, J. M., Ramalho, E. A. V. F., Melo, T. V. C., Silva, I. S. M., Júnior, G. S. Lins, Bastos, M. L. A., Farias, D. F., Souza, P. C. V., Carvalho, A. F. U., Silva, M. C. S., Almeida, R. M. R. G., and E. Neto, R., “Genetic variability in the ITS and IGS regions of the ribosomal DNA of Acremonium cavaraeanum exhibiting antimicrobial activity”, vol. 12, pp. 6983-6995, 2013.

Endophytic microorganisms represent promising alternatives for obtaining new drugs of biotechnological importance. In this study, the endophytic species Acremonium cavaraeanum (A1a) isolated from Cocos nucifera was cultivated for the production of secondary metabolites, and its extracts and fractions were evaluated by the dilution method (MIC). The EtOAc extracts and MeOH fractions were tested against Gram-positive and -negative bacteria, and had an MIC of 125 μg/mL when evaluated in the EtOAc extract (EBI).

Optimizing the efficiency of the touchdown technique for detecting inter-simple sequence repeat markers in corn (Zea mays)

E. C. Oliveira, Júnior, A. T. Amaral, Gonçalves, L. S. A., Pena, G. F., Júnior, S. P. Freitas, Ribeiro, R. M., and Pereira, M. G., “Optimizing the efficiency of the touchdown technique for detecting inter-simple sequence repeat markers in corn (Zea mays)”, vol. 9, pp. 835-842, 2010.

We evaluated the efficiency of the touchdown method to determine the ideal PCR conditions for distinct inter-simple sequence repeat primers for processing DNA from common corn, popcorn, sweet corn, and a Tripsacum-maize hybrid. Genomic DNA was extracted from eight accessions of corn: two of the dent type, one Tripsacum-maize hybrid, one sweet corn, one flint-type corn, and three popcorn.

M13-microsatellite PCR and rDNA sequence markers for identification of Trichoderma (Hypocreaceae) species in Saudi Arabian soil

K. A. Abd-Elsalam, Almohimeed, I., Moslem, M. A., and Bahkali, A. H., “M13-microsatellite PCR and rDNA sequence markers for identification of Trichoderma (Hypocreaceae) species in Saudi Arabian soil”, vol. 9, pp. 2016-2024, 2010.

Seven fungal isolates were identified as pan-global Hypocrea/Trichoderma species, from section Trichoderma, on the basis of their morphology. These species were H. lixii/T. harzianum and H. orientalis/T. longibrachiatum. PCR-based markers with primer M13 (core sequence of phage M13) and internal-transcribed spacer sequences of ribosomal DNA were used to confirm the identity of the two Trichoderma species.