Post-injection hybridization of complementary DNA strands on capillary electrophoresis platforms: A novel solution for dsDNA artifacts

Several laboratories have reported the occurrence of a split or n − 1 peak at the vWA locus in PowerPlex^® 16 and PowerPlex^® ES amplification products separated on 4- and 16-capillary electrophoresis instruments. The root cause of this artifact is post-PCR reannealing of the unlabeled, unincorporated vWA primer to the 3′-end of the tetramethylrhodamine (TMR)-labeled strand of the vWA amplicon. This reannealing occurs in the capillary post-electrokinetic injection. The split peak is eliminated by incorporation into the loading cocktail of a sacrificial hybridization sequence (SHS) oligonucleotide that is complementary to the vWA primer. The SHS preferentially anneals to the primer instead of the TMR-labeled strand of the vWA amplicon. In addition, the n − 10/n − 18 artifact that may be seen at the vWA locus was determined to be due to double-stranded amplicon formed post-electrokinetic injection into the capillary. This was also eliminated by adding in two Complementary Oligo Targets (COT1 and COT2) in addition to the SHS oligonucleotide into the loading cocktail. These three oligonucleotides are complementary to the 33 bases at the 5′-end of the unlabeled vWA amplicon strand and the 60 bases at its 3′-end and therefore compete for hybridization to the TMR-labeled amplicon strand. Incorporation of these three oligonucleotides in the Internal Lane Standard 600 (ILS600) eliminate both the split peak and n − 10/n − 18 artifact in PowerPlex^® 16 and PowerPlex^® ES amplification products without affecting sizing of alleles at the vWA locus or any locus in the PowerPlex^® 16, PowerPlex^® Y, PowerPlex^® ES, AmpFlSTR^® Profiler Plus^® ID, AmpFlSTR^® Cofiler^®, and AmpFlSTR^® SGM Plus^® kits.     

Business Routes of Influence in Brussels: Exploring the Choice of Direct Representation

This paper presents one of the first statistical analyses of the factors that determine the extent to which direct influence strategies are used by companies. New survey evidence and multivariate analysis is used to assess seven hypotheses to explain direct influence strategies. The factor of greatest significance is company size since direct influence is open only to large companies. But the extent to which direct influence is followed by large companies depends on their sector, its extent of organization, the sector's size and 'weight', and the form of the business association relevant to the sector. Direct influence is more likely where associations cover large corporate businesses (rather than owner-managers, professionals, federations or mixed associations), in sectors with a high degree of market concentration within large companies, and where their associations are relatively small. These findings are related to the logic of collective action suggesting that in sectors where there is the least chance to opt out of an association, there is a greater chance of a business also directly lobbying in order to assure itself that its interests are not being diluted and so that it can gain direct specific information or influence benefits.     

Zum grundbücherlichen Richtigkeitsgebot und seiner amtswegigen Verwirklichung (§ 28 LiegTeilG) (1. Teil)

Die übereinstimmung zwischen Grundbuch, Kataster und Realit?t liegt im Schnittpunkt zwischen Gesetzgebung und ?konomie im ?ffentlichen Interesse. Im LiegTeilG, sohin an unvermuteter Stelle, ist ein Verfahren zur amtswegigen Verwirklichung dieses Kongruenzprinzips normiert, dessen prozessuales Instrumentarium durch auch jüngste Gesetzgebungsakte direkt und indirekt deformiert worden ist. Diese Abhandlung untersucht, ob die ursprünglich verfolgten gesetzgeberischen Intentionen zur Herstellung der Grundbuchsordnung auf der geltenden Rechtslage im judikativen Weg gem §§ 6 f ABGB revitalisierbar sind oder ob es zu deren Reanimation eines legistischen Eingriffs bedarf.     

Five Words for Assisted Dying

Motivated by Lord Joffe’s Assisted Dying for the Terminally Ill Bill, but with one eye on any possible future legislation, I consider the justifications that might be offered for limiting assistance in dying to those who are suffering unbearably from terminal illness. I argue that the terminal illness criterion and the unbearable suffering criterion are not morally defensible separately: that a person need be neither terminally ill (or ill at all), nor suffering unbearably (or suffering at all) to have a right to assisted dying. Indeed: I shall suggest that the unbearable suffering criterion undermines the Bill (or any proposal like it) wholesale. On the other hand, the criteria taken together are defensible, and this defence would be built on a concern for the protection of the vulnerable. However, I also claim that this implies that the law might justifiably—and maybe even properly—aim to prevent a person from gaining access to that to which they have a serious moral right. This seems paradoxical, and, towards the end of the paper, I seek to tease apart the paradox.     

The Global Era of National Policy and the Pan-Oceanic National Fleet

This article develops a framework for considering the future requirements for U.S. maritime power. It does so by proposing that these requirements must be considered within the context of an integrated National Fleet—the combined capabilities of the three “Sea Services”—the U.S. Navy, U.S. Marine Corps, and U.S. Coast Guard. It also would include their reserve components, as well as the Military Sealift Command (MSC) and the Maritime Administration's Ready Reserve Force (RRF)—the two organizations responsible for maintaining the nation's strategic sealift fleet.