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.     

A response to charles t. goodsell

Goodsell's definition of rituals and administration logically and experientially rings true. Formalizing the relationship between them was both unique and obvious, since we all have an intuitive sense that some of our administrative habits are ritualistic; we regularly complete paperwork, stop for coffee, make phone calls, etc. However, Goodsell has afforded us the opportunity to refer to some of these habits, such as red tape (immigration procedures) or administrivia (staff meetings) as rituals which help promote the social continuity. Similarly, we can rationalize that our comfortable administrative habits need not be changed because they unify.

Yes, the concept of rituals can be applied to many aspects of administration, but to what end? Ritualists suggest some rituals “work” while others do not, so we know little about when or how rituals are effective. To simply state that the concepts and philosophies of rituals offer a means to better understand administration leaves one cold. Goodsell provides little help in discerning how and why rituals are important to understanding administration. Since he links administration to rituals without an explicit purpose for doing so, he contributes to the perception that bureaucracies consist of meaningless tasks and tangles that prevent efficient government operations.     

Reparations in the Long Run

This paper explores how policy-makers during the Second World War attempted to “learn the lessons” of history from the reparations settlement imposed after the First World War. It shows how these lessons were developed and articulated in the formulation of, in particular, American foreign policy, and also their consequences for foreign policy during and after the Second World War. The paper demonstrates the important role of European advisors in shaping American policy, thereby illustrating that not all American lessons of history were born in the USA. It also draws out how many of these lessons have found an echo in the historiography of German reparations that has emerged over the past fifty years. In both periods the issues of enforcement and compliance, the issues that concern us generally in this volume, dominated the debate between advisors and policy-makers.     

A Rapid and Efficient Method for Evaluation of Suspect Testimony: Palynological Scanning

A rapid method for evaluating suspect testimony is valuable at any stage in an inquiry and can result in a change of direction in an investigation. Rape cases, in particular, can present problems where a defendant renders DNA analysis redundant by claiming that the claimant consented to have sexual relations. Forensic palynology is valuable in confirming or eliminating locations as being crime scenes, thus checking the testimony of both parties. In contrast to some forensic disciplines, forensic palynology can provide critical information without time-consuming full analysis. Two cases are described where the palynological assemblages from comparator samples of pertinent places were compared with those obtained from clothing of claimants and defendants. The results of rapid microscopical scanning of relevant preparations led to early confessions, thus obviating the need for costly analyses and protracted court proceedings. A third case demonstrates the unbiased nature of this technique where a man, although innocent of any offense, lied about having visited the crime scene for fear of prosecution. This highlights the need for sensitive policing in claims of rape.