RT-Bst: an integrated approach for reverse transcription and enrichment of cDNA from viral RNA

The synthesis of cDNA from RNA is challenging due to the inefficiency of reverse transcription (RT). In order to address this, a method was developed known as RT-Bst for sequential RT of RNA and Bst DNA polymerase amplification for enrichment of cDNA in a single tube reaction. Using genomic RNA from bacteriophage MS2, the yield of cDNA produced by RT alone and RT-Bst were compared by analysis of PCR-amplified products. Using random primers a superior performance was observed when amplifying MS2 RNA following RT-Bst compared to RT alone, indicating that greater quantities of cDNA were present after RT-Bst. RT-Bst was also compared with RT alone for their relative ability to produce sufficient cDNA to amplify 8 target regions spanning the respiratory syncytial virus (RSV) genome. Six out of 8 targets were amplified consistently by PCR subsequent to RT-Bst amplification whereas only 3 out of 8 targets could be amplified after RT alone. RSV sequences were selectively amplified using RSV specific primers from a mixed template containing an excess of MS2 RNA in a RT-Bst reaction without amplifying MS2 sequences. This suggests that RT-Bst can be used to amplify RNA sequences non-specifically using random primers and specifically using sequence specific primers and enhances the yield of cDNA when compared to RT alone.

Assessing energy consumption in supply chains

The paper addresses the transport activities and associated energy consumption involved in the production and supply of two products: jeans and yoghurt. In the case of jeans, the analysis is from the locations in which cotton is grown, to retail outlets in the UK; in the case of yoghurt, the analysis is from the supply of milk on farms, to retail outlets in France. The results show that the transport stages from the point of jeans manufacture to UK port are responsible for the greatest proportion of transport energy use per kilogram of jeans in the UK supply chain. In the case of the French yoghurt supply chains, the results indicate that each of the three transport stages from farm to third-party distribution centre consume approximately the same proportion of total freight transport energy. The energy used on the transport stage for yoghurt from third-party distribution centre to retail outlet varies depending on the type of retail outlet served. Far greater quantities of energy are used in transporting jeans than yoghurts from farm/field to retail outlet. This is explained by the distances involved in the respective supply chains. Both case studies demonstrate that the energy used by consumers transporting goods to their homes by car can be as great as total freight transport energy used in the supply chain from farm/field to retail outlet (per kilogram of product transported).

Investigating relationships between road freight transport, facility location, logistics management and urban form

This paper examines road freight transport activity and its relationship with facility location, logistics management and urban form through an analysis of 14 selected urban areas in the UK. Improved understanding of this relationship will assist planners when making transport and land use decisions. The findings suggest that several geographical, spatial and land use factors have important influences on freight activity in urban areas. Commercial and industrial land use patterns affect the types and quantities of goods produced, consumed, and hence the total quantity of freight transport handled. This also influences the distances over which goods are moved and by what specific mode. There has been relatively low growth in warehousing in many of the selected areas over the last decade compared to the national average as well suburbanisation of warehousing in some locations. This affects the origin and destination of journeys visiting these facilities and typically increases the distance of such journeys. A greater proportion of road freight has been shown to be lifted on internal journeys in large urban areas than in smaller ones. Journeys within urban areas have been shown to be less efficient than journeys to and from the urban area in the 14 locations studied due to the much smaller average vehicle carrying capacities and lower lading factors for journeys within urban areas. The length of haul on journeys to and from urban areas studied was found to be greatest for those areas with a major seaport and/or which were geographically remote. This affects the road freight transport intensity of goods transport journeys.