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.

Smart City-Regional Governance: A 'Dual Transition'

This paper introduces the concept of ‘dual transition’ in relation to the notion of ‘smartness’ in city-regional governance. This consists of two intersecting dimensions, a broader change in political-economic and societal circumstances which surround city-regions, and a more detailed, local change towards ‘smartness’ in policy practices. Although ‘smart’ seems to have become a somewhat ubiquitous adjective in urban policy, it seeks to project policies that seek to go beyond a one-dimensional ‘growth agenda’ by addressing also the multifaceted quest for social, economic and environmental sustainability. The underlying shift in discourse, rationality and suggested policy responses in both dimensions may be captured by the concept of ‘transition’ in relation to policies and governance. Originally developed in conjunction with broad political-economic regime change, such as post-authoritarian democratisation, the idea of ‘transition’ has also been applied to the more specific concept of sustainability as ‘sustainability transition’.