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.     

Intoxicated Consent in Rape: Bree and Juror Decision-Making

The article examines the recent Court of Appeal judgment in Bree, in which the court considered the guidance that should be given to a jury when the complainant is intoxicated at the time of an alleged rape. As well as analysing the reasoning of the court in Bree, the judgment is placed in context by examining factors, such as social attitudes, that influence juror decision-making in rape cases. It also examines the suggestion made by the court in Bree that further law reform in this area would represent 'patronising interference' in the lives of women.     

Left realist criminology: Strengths,weaknesses and the feminist critique

Although there is an already large British literature both supporting and attacking left realism, and a growing North American interest on the subject among criminologists, there has been surprisingly little written which attempts to locate both the strengths and weaknesses of the left realist position on crime control. Perhaps the place where the left realists may be weakest is in response to a feminist critique. Actually, it is not only left realism but the socialist left in general which has been unsuccessful in providing adequate responses to the issues brought forth by feminists. This paper attempts to locate the position of left realism within the left criminology debate, and to find its strong and weak points. Further, it attempts to explicate the feminist critique, and to suggest responses critical criminologists might explore, such as those proposed by peacemaking criminology. *** DIRECT SUPPORT *** AW502012 00004     

Interpretation of unusual wounds caused by firearms.

The interpretation of the range of fire of gunshot wounds requires coordination of information and observations from the autopsy surgeon, scene investigator, and laboratory analyst. Opinions based on incomplete information often lead to misinterpretation of the wound. The effects of interposed targets such as clothing, windows, and body parts are discussed. Case examples are given for interposed targets that cause confusing patterns and even lead to misidentification of the points of entrance and exit of the projectile in the body. The splitting of tissue in areas of bony prominence can be mistaken for evidence of discharge of a firearm at close range.     

HLA DQ alpha typing of forensic specimens by amplification restriction fragment polymorphism (ARFP) analysis.

The alleles present at the HLA DQ alpha locus may be typed by either allele specific oligonucleotide (ASO) probing or by restriction mapping, referred to here as amplification restriction fragment polymorphism (ARFP) analysis. ASO typing relies upon hybridization principles, whereas ARFP typing relies upon restriction site analysis. Dot-blot ASO typings which are of doubtful interpretation may be directly checked by ARFP analysis. Aliquots of the PCR products amplified using the commercial Amplitype HLA DQ alpha system are digested with suitable restriction endonucleases. Electrophoresis, blotting and detection of biotin-labelled restriction fragments provides a sensitive and robust typing method suited to forensic analysis.     

Family violence and Asian drinkers

This is a study of 70 consecutive referrals to the Department of Psychological Medicine, National University Hospital, for treatment of drinking problems. There were 31 Chinese and 39 Indian drinkers, with 63 males and 7 females. Family violence with physical abuse occurred in 21 cases (30%). One of three wives of drinkers had been physically abused, but no children were victims. Drinkers with family violence were generally younger and more severely dependent on alcohol than the non-violent drinkers. There was no relationship between family violence and the ethnicity or marital status of the drinkers.