We have developed a new method for identifying specific single- or double-stranded DNA sequences called nicking endonuclease signal amplification (NESA). A probe and target DNA anneal to create a restriction site that is recognized by a strand-specific endonuclease that cleaves the probe into two pieces leaving the target DNA intact. The target DNA can then act as a template for fresh probe and the process of hybridization, cleavage and dissociation repeats. Laser-induced fluorescence coupled with capillary electrophoresis was used to measure the probe cleavage products. The reaction is rapid; full cleavage of probe occurs within one minute under ideal conditions. The reaction is specific since it requires complete complementarity between the oligonucleotide and the template at the restriction site and sufficient complementarity overall to allow hybridization. We show that both Bacillus subtilis and B. anthracis genomic DNA can be detected and specifically differentiated from DNA of other Bacillus species. When combined with multiple displacement amplification, detection of a single copy target from less than 30 cfu is possible. This method should be applicable whenever there is a requirement to detect a specific DNA sequence. Other applications include SNP analysis and genotyping. The reaction is inherently simple to multiplex and is amenable to automation.
INTRODUCTION: Student hardship in social work has become the subject of research in recent years. Social work students face particular challenges because of the financial, social and emotional demands of long, unpaid, clinical placements. APPROACH: This article reports on a literature review conducted prior to a mixed-methods study commenced in 2019. This review informed the development of a survey of social work students and recent graduates and a set of qualitative interviews that will be reported elsewhere. A further review was conducted in July 2021 to inform the analysis. FINDINGS: A recurring theme throughout much of the reviewed social work literature examined for this project has been the intensity of struggle that students face in their study. Across the reviewed literature, students, educators, and researchers suggest interventions to reduce hardship. Gaps remain in our understanding of the intersectional factors of ethnicity, gender and disability and, in particular, the impact on Aotearoa New Zealand students.
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