A detailed protocol for subcellular RNA sequencing (subRNA-seq)

In eukaryotic cells, RNAs at various maturation and processing levels are distributed across cellular compartments. The standard approach to determine transcript abundance and identity in vivo is RNA sequencing (RNA-seq). RNA-seq relies on RNA isolation from whole-cell lysates and thus mainly captures fully processed, stable, and more abundant cytoplasmic RNAs over nascent, unstable, and nuclear RNAs. Here, we provide a step-by-step protocol for subcellular RNA-seq (subRNA-seq). subRNA-seq allows the quantitative measurement of RNA polymerase II-generated RNAs from the chromatin, nucleoplasm, and cytoplasm of mammalian cells. This approach relies on cell fractionation prior to RNA isolation and sequencing library preparation. High-throughput sequencing of the subcellular RNAs can then be used to reveal the identity, abundance, and subcellular distribution of transcripts, thus providing insights into RNA processing and maturation. Deep sequencing of the chromatin-associated RNAs further offers the opportunity to study nascent RNAs. Subcellular RNA-seq libraries are obtained within 5 day

Evaluating the character and preservation of DNA within allophane clusters in buried soils on Holocene tephras, northern New Zealand

Clay minerals possess sorptive capacities for organic and inorganic matter, including DNA (Lorenz and Wackernagel, 1994), and hence reduce the utilization and degradation of organic matter or DNA by microorganisms. Buried allophane-rich soils on tephras (volcanic-ash beds) on the North Island, dated using tephrochronology, provide a valuable paleobiological ‘laboratory’ for studying the preservation of ancient DNA (aDNA) (Haile et al., 2007). Allophane comprises Al-rich nanocrystalline spherules ~3.5-5 nm in diameter (Fig. 1) with extremely large surface areas (up to 1000 m2 g-1). Moreover, allophanic soils are strongly associated with organic matter (Parfitt, 2009), and so we hypothesize that allophane also plays an important role for DNA protection within such soils

A holistic multi-methodology for sustainable renovation

A review of the barriers for building renovation has revealed a lack of methodologies, which can promote sustainability objectives and assist various stakeholders during the design stage of building renovation/retrofitting projects. The purpose of this paper is to develop a Holistic Multi-methodology for Sustainable Renovation, which aims to deal with complexity of renovation projects. It provides a framework through which to involve the different stakeholders in the design process to improve group learning and group decision-making, and hence make the building renovation design process more robust and efficient. Therefore, the paper discusses the essence of multifaceted barriers in building renovation regarding cultural changes and technological/physical changes. The outcome is a proposal for a multi-methodology framework, which is developed by introducing, evaluating and mixing methods from Soft Systems Methodologies (SSM) with Multiple Criteria Decision Making (MCDM). The potential of applying the proposed methodology in renovation projects is demonstrated through a case study

Russell Ackoff

Russell Ackoff (usually known as ‘Russ’) was a pioneer of the application of systems approaches to management, both through theoretical developments and through a deep and practical engagement with many different organisations. He was a strong advocate of the need for systems approaches to take full account of the complexity of inter-related problems and not simply to present glib technical solutions

Mediating boundaries between knowledge and knowing: ICT and R4D praxis

Research for development (R4D) praxis (theory-informed practical action) can be underpinned by the use of Information and Communication Technologies (ICTs) which, it is claimed, provide opportunities for knowledge working and sharing. Such a framing implicitly or explicitly constructs a boundary around knowledge as reified, or commodified – or at least able to be stabilized for a period of time (first order knowledge). In contrast ‘third-generation knowledge’ emphasizes the social nature of learning and knowledge-making; this reframes knowledge as a negotiated social practice, thus constructing a different system boundary. This paper offers critical reflections on the use of a wiki as a data repository and mediating technical platform as part of innovating in R4D praxis. A sustainable social learning process was sought that fostered an emergent community of practice among biophysical and social researchers acting for the first time as R4D co-researchers. Over time the technologically mediated element of the learning system was judged to have failed. This inquiry asks: How can learning system design cultivate learning opportunities and respond to learning challenges in an online environment to support R4D practice? Confining critical reflection to the online learning experience alone ignores the wider context in which knowledge work took place; therefore the institutional setting is also considered

Not in my back yard! Sports stadia location and the property market

In recent years sports stadia have been built in the UK, not only for their intended sporting purpose but with the twin aim of stimulating economic and physical regeneration. However, proposals to locate stadia in urban areas often prompt a negative reaction from local communities, fearing a decline in property prices. This paper will use a case study of the Millennium Stadium in Cardiff and the City of Manchester Stadium to illustrate that in contrast to this widely held belief, sports stadia can actually enhance the value of residential property. Furthermore, it will argue that stadia also contribute indirectly to property value through the creation of pride, confidence and enhanced image of an area.</p

Soft systems methodology: a context within a 50-year retrospective of OR/MS

Soft systems methodology (SSM) has been used in the practice of operations research and management science OR/MS) since the early 1970s. In the 1990s, it emerged as a viable academic discipline. Unfortunately, its proponents consider SSM and traditional systems thinking to be mutually exclusive. Despite the differences claimed by SSM proponents between the two, they have been complementary. An extensive sampling of the OR/MS literature over its entire lifetime demonstrates the richness with which the non-SSM literature has been addressing the very same issues as does SSM