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Int J Legal Med (2012) 126:601606 DOI 10.1007/s00414-012-0706-6
ORIGINAL ARTICLE
The validation of a 15 STR multiplex PCR for Cannabis species
Stephan Khnemann & Johanna Nedele &
Daniela Schwotzer & Julia Morzfeld & Heidi Pfeiffer
Received: 31 January 2012 /Accepted: 26 April 2012 /Published online: 10 May 2012 # Springer-Verlag 2012
Abstract Trade and acquisition of Cannabis drugs are illegal in many countries worldwide; nevertheless, crimes related with these drugs are a major problem for the investigative authorities. With this manuscript, we want to introduce a 15 short tandem repeat (STR) Cannabis marker set that can be amplified in one PCR reaction. This multiplex PCR is specific to Cannabis species and combines highly informative STR markers. The 15 STR multiplex is easy to use and was validated according to common laboratory quality standards. Due to the fact that a lot of Cannabis plants are cultivated by clonal propagation and may show aneuploidy, polyploidy or multiple gene loci, it is not possible to apply biostatistics that follow the HardyWeinberg law. However, this multiplex will help the police to trace back trade routes of drug syndicates or dealers and it can help to link Cannabis plants to a crime scene.
Keywords Cannabis DNA short tandem repeat . STR
Introduction
Cannabis species are economic plants used for the production of food, fibres, oils and intoxicants [1]. Some of them belong to the most frequently used illicit drugs worldwide
and are therefore prohibited by law in many countries [2]. The main psychoactive compound of drug Cannabis is 9-tetrahydrocannabinol (THC) which can be found in high concentrations in leaves and inflorescences.
Techniques to distinguish Cannabis on the DNA level were developed previously by different working groups [37]. Short tandem repeats (STRs) appear to have the best discrimination ability; nevertheless, none of these groups tried to combine more than six STRs in one multiplex PCR. STRs are repetitive sequences of up to six bases at a defined gene locus, found normally in the non-coding region of autosomal or gonosomal DNA. They should be flanked by a conserved DNA region so that it is possible to create homologues and species-specific primers for PCR amplification.
The idea of using DNA analysis on Cannabis was first triggered by the question of whether or...