DNA isolation

Comparison of methods to isolate DNA from Caesalpinia ferrea

C. C. Sousa, Gomes, S. O., Lopes, A. C. A., Gomes, R. L. F., Britto, F. B., Lima, P. S. C., and Valente, S. E. S., “Comparison of methods to isolate DNA from Caesalpinia ferrea”, vol. 13. pp. 4486-4493, 2014.

Molecular markers are important for characterizing the genetic diversity of plants and can provide the basis for strategies to protect and conserve endangered populations. However, numerous molecular techniques are used, requiring an evaluation of fast and efficient methods to extract DNA. Since molecular studies of Caesalpinia ferrea are rare, it is important to develop and/or adapt a DNA extraction protocol that produces quality DNA samples to enable the design of strategies for the conservation of this threatened species.

An economical and combined method for rapid and efficient isolation of fungal DNA

T. Lech, Syguła-Cholewinska, J., and Szostak-Kot, J., “An economical and combined method for rapid and efficient isolation of fungal DNA”, vol. 13, pp. 10779-10786, 2014.

DNA isolation is a crucial step of conducting genetic studies in any organism. However, this process is quite difficult when studying fungi because of the need to damage the fungal cell walls of specific structures. In this study, we developed a method for the rapid and efficient isolation of fungal DNA based on simultaneous mechanical and enzymatic cell wall degradation. There are several typical modifications of the standard phenol-chloroform DNA extraction method. This method can be modified to degrade the fungal cell wall.

Simple and rapid protocol for the isolation of PCR-amplifiable DNA from medicinal plants

A. A. Alatar, Mahmoud, M. A., Al-Sohaibani, S. A., and Abd-Elsalam, K. A., “Simple and rapid protocol for the isolation of PCR-amplifiable DNA from medicinal plants”, vol. 11, pp. 348-354, 2012.

Medicinal plant species has a valuable economic importance because of its usage as pharmaceuticals, nutritional, as well as its use in popular medication. For DNA-based techniques, nanogram quantities of the purified DNA are requisite to amplify and yield sufficient amounts of PCR products. SDS-based DNA isolation method was used to extract DNA from 11 species of different aromatic and medicinal plants collected from Saudi Arabia. Three hundred milligrams of fresh shredded plant material was necessary.

DNA extraction from skins of wild (Hydrochoerus hydrochaeris and Pecari tajacu) and domestic (Sus scrofa domestica) species using a novel protocol

G. N. Ojeda, Amavet, P. S., Rueda, E. C., and Siroski, P. A., “DNA extraction from skins of wild (Hydrochoerus hydrochaeris and Pecari tajacu) and domestic (Sus scrofa domestica) species using a novel protocol”, vol. 11, pp. 672-678, 2012.

Sometimes, commercial products obtained from wild animals are sold as if they were from domestic animals and vice versa. At this point of the productive chain, legal control of possible wildlife products is difficult. Common in the commerce of northern Argentina, skins of two wild species, the carpincho and the collared peccary, look very similar to each other and to those of the domestic pig; it is extremely difficult to differentiate them after they have been tanned.

Identification of the isoamylase 3 gene in common wheat and its expression profile during the grain-filling period

G. Z. Kang, Liu, G. Q., Xu, W., Zhu, Y. J., Wang, C. Y., Ling, H. Q., and Guo, T. C., “Identification of the isoamylase 3 gene in common wheat and its expression profile during the grain-filling period”, vol. 12, pp. 4264-4275, 2013.

In higher plants, isoamylase-type starch debranching enzyme catalyzes the α-1,6-glucosidic linkages of glycogen and phytoglycogen. We cloned an isoamylase-type starch debranching enzyme ISA3 cDNA sequence (2883 bp), designated as TaISA3, from common wheat (Triticum aestivum), using the rapid amplification of cDNA ends method. The open reading frame of TaISA3 was found to have 2331 bp, and its deduced amino acid sequence was found to share high similarity with those of other gramineous plant ISA3 proteins.

An efficient protocol for tissue sampling and DNA isolation from the stem bark of Leguminosae trees

R. M. L. Novaes, Rodrigues, J. G., and Lovato, M. B., “An efficient protocol for tissue sampling and DNA isolation from the stem bark of Leguminosae trees”, vol. 8, pp. 86-96, 2009.

Traditionally, molecular studies of plant species have used leaves as the source of DNA. However, sampling leaves from tall tree species can be quite difficult and expensive. We developed a sequence of procedures for using stem bark as a source of DNA from Leguminosae trees of the Atlantic Forest and the Cerrado. Leguminosae is an important species-rich family in these two highly diverse and endangered biomes. A modified CTAB protocol for DNA isolation is described, and details of the procedures for sampling and storage of the bark are given.

CENP-B is a conserved gene among vegetal

O. Barbosa-Cisneros and Herrera-Esparza, R., “CENP-B is a conserved gene among vegetal”, vol. 1, pp. 241-245, 2002.

To explore the CENP-B centromere protein in beans, carrots, onions and potatoes, total RNA was isolated and reverse transcribed by PCR, and the cDNA encoding the CENP-B amino terminus domain amplified using CENP-B oligonucleotides. Blots containing PCR products were hybridized with a nick-translated pG/CNPB probe containing a complete human CENP-B gene. In all the plant species, anti-CENP-B antibodies recognized an 80-kDa protein.

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