Multiplex short tandem repeat amplification of low template DNA samples with the addition of proofreading enzymes

Abstract:  With <100 pg of template DNA, routine short tandem repeat (STR) analysis often fails, resulting in no or partial profiles and increased stochastic effects. To overcome this, some have investigated preamplification methods that include the addition of proofreading enzymes to the PCR cocktail. This project sought to determine whether adding proofreading polymerases directly in the STR amplification mixture would improve the reaction when little template DNA is available. Platinum Taq High Fidelity and GeneAmp High Fidelity were tested in Profiler Plus? STR reactions alone and in combination with AmpliTaq^® Gold. All reactions included the additional step of a post‐PCR purification step. With both pristine low template DNA and casework samples, the addition of these polymerases resulted in comparable or no improvement in the STR amplification signal. Further, stochastic effects and artifacts were observed equally across all enzyme conditions. Based on these studies, the addition of these proofreading enzymes to a multiplex STR amplification is not recommended for low template DNA work.     

LESSENING THE STING OF ASFA: THE REHABILITATION‐RELAPSE DILEMMA BROUGHT ABOUT BY DRUG ADDICTION AND TERMINATION OF PARENTAL RIGHTS

Congress passed the Adoption and Safe Families Act of 1997 (ASFA) as a response to children waiting in foster homes for years without permanent placement. In addressing the problem of permanency, however, Congress set a strict limit on how long a child could be in foster care (15 out of the most recent 22 months) before a state must either commence a proceeding to terminate parental rights or else lose valuable federal funding. Due to health care funding schemes and quality of treatment, this requirement, in particular, negatively impacts parents currently in drug rehabilitation whose parental rights may be permanently terminated before a realistic chance to recover is permitted. Although ASFA requires that states make “reasonable efforts” to keep families united, it does not define “reasonable efforts,” leaving parental rights and family unity subject to a chaotic interpretation of this requirement from state to state. “Reasonable efforts” should be interpreted to take into account current drug addiction and recovery research and drug court programs should be used to facilitate this goal. Research has shown that focusing on adequate treatment saves states money and improves the lives of children and their families, reducing the need for reliance on termination of parental rights.