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 Judicial‐Mental Health Partnership to Heal Young Children in Juvenile Court

In this article, we describe the background and issues to be addressed related to dependent children in juvenile court. In an important effort to systematically examine developmental functioning and treatment needs in maltreated and violence‐exposed young children, the Prevention and Evaluation of Early Neglect and Trauma (PREVENT) initiative of the Dependency Court Intervention Program for Family Violence, a national demonstration project in the Miami‐Dade Juvenile Court, developed a program to evaluate all infants, toddlers, and preschoolers who are adjudicated dependent by the court. The goal of the intervention is to raise awareness of the needs of infants and toddlers in juvenile court and to work toward healing the child. The PREVENT program involved the evolution of a judicial‐mental health partnership designed to assist the court in making more informed decisions about the best interest of the child by adding scientific knowledge about development, prevention, intervention, evaluation, and treatment. The outcome of the partnership and multidisciplinary approach is illustrated through presenting a case vignette of a mother and baby showing the challenges and strengths of intervention. Finally, we consider overall outcomes of the intervention and directions for the future.     

Phosphoglucomutase (PGM) grouping of bloodstains on silver.

This paper describes a case involving alteration of phosphoglucomutase (PGM) isoenzyme patterns in bloodstains present on silver. The effect could be produced by treating blood samples with silver nitrate solution.     

PROFESSIONAL INTERDEPENDENCE IN DIVORCE PRACTICES

Divorce mediation is one area of the new and growing field of interpersonal mediation. Differences and similarities between divorce mediation and divorce psychotherapy are discussed in this article, and a method for teamwork between the two professions is proposed. The authors take the approach that the separate but overlapping practices of mediation and psychotherapy can be interdependent and that specific benefits to divorcing clients can result when divorce mediation and divorce psychotherapy are practiced collaboratively.