Big data from small tissue extraction of high-quality RNA for RNA-Sequencing from different oilseed Brassica seed tissues during seed development

Background Obtaining high-quality RNA for gene expression analyses from different seed tissues is challenging due to the presence of various contaminants, such as polyphenols, polysaccharides and lipids which interfere with RNA extraction methods. At present, the available protocols for extracting RNA from seeds require high amounts of tissue and are mainly focused on extracting RNA from whole seeds. However, extracting RNA at the tissue level enables more detailed studies regarding tissue specific transcriptomes during seed development. Results Seeds from heart stage embryo to mature developmental stages of Brassica napus and B. oleracea were sampled for isolation of the embryo, endosperm and seed coat tissues. Ovules and ovary wall tissue were also collected from pre-fertilized buds. Subsequent to testing several RNA extraction methods, we determined that modifications applied to E.Z.N.A. Plant RNA and Picopure RNA Isolation kit extraction methods resulted in RNA with high yield and quality. Furthermore, the use of polyvinylpolypyrrolidone for the extraction of RNA from seed coats and endosperm at green seed stage resulted in high-quality RNA. As a result of the introduced modifications to established RNA extraction methods, the RNA from all the above-mentioned tissues presented clear 28S and 18S bands and high RIN values, ranging from 7.0 to 10.0. The protocols reported in this study are not only suitable for different and challenging seed tissue types, but also enable the extraction of high-quality RNA using only 2 to 3 mg of starting tissue. Conclusions Here, we present efficient, reproducible and reliable high-quality RNA extraction methods for diverse oilseed Brassica spp reproductive tissue types including pre-fertilization and developing seed tissues for diploid and polyploid species. The high-quality RNA obtained is suitable for RNA-Sequencing and subsequent gene expression analysis

Quantum Mechanics at Planck's scale and Density Matrix

In this paper Quantum Mechanics with Fundamental Length is chosen as Quantum Mechanics at Planck's scale. This is possible due to the presence in the theory of General Uncertainty Relations. Here Quantum Mechanics with Fundamental Length is obtained as a deformation of Quantum Mechanics. The distinguishing feature of the proposed approach in comparison with previous ones, lies on the fact that here density matrix subjects to deformation whereas so far commutators have been deformed. The density matrix obtained by deformation of quantum-mechanical density one is named throughout this paper density pro-matrix. Within our approach two main features of Quantum Mechanics are conserved: the probabilistic interpretation of the theory and the well-known measuring procedure corresponding to that interpretation. The proposed approach allows to describe dynamics. In particular, the explicit form of deformed Liouville's equation and the deformed Shr\"odinger's picture are given. Some implications of obtained results are discussed. In particular, the problem of singularity, the hypothesis of cosmic censorship, a possible improvement of the definition of statistical entropy and the problem of information loss in black holes are considered. It is shown that obtained results allow to deduce in a simple and natural way the Bekenstein-Hawking's formula for black hole entropy in semiclassical approximation.Comment: 18 pages,Latex,new reference

Bromination of Graphene and Graphite

We present a density functional theory study of low density bromination of graphene and graphite, finding significantly different behaviour in these two materials. On graphene we find a new Br2 form where the molecule sits perpendicular to the graphene sheet with an extremely strong molecular dipole. The resultant Br+-Br- has an empty pz-orbital located in the graphene electronic pi-cloud. Bromination opens a small (86meV) band gap and strongly dopes the graphene. In contrast, in graphite we find Br2 is most stable parallel to the carbon layers with a slightly weaker associated charge transfer and no molecular dipole. We identify a minimum stable Br2 concentration in graphite, finding low density bromination to be endothermic. Graphene may be a useful substrate for stabilising normally unstable transient molecular states

Resilience and well-being among children of migrant parents in South-East Asia

There has been little systematic empirical research on the well-being of children in transnational households in South-East Asia—a major sending region for contract migrants. This study uses survey data collected in 2008 from children aged 9, 10 and 11 and their caregivers in Indonesia, the Philippines, and Vietnam (N=1,498). Results indicate that while children of migrant parents, especially migrant mothers, are less likely to be happy compared to children in non-migrant households, greater resilience in child well-being is associated with longer durations of maternal absence. There is no evidence for a direct parental migration effect on school enjoyment and performance. The analyses highlight the sensitivity of results to the dimension of child well-being measured and who makes the assessment.Publisher PDFPeer reviewe