Extraction of high quality DNA from seized Moroccan cannabis resin (Hashish).

The extraction and purification of nucleic acids is the first step in most molecular biology analysis techniques. The objective of this work is to obtain highly purified nucleic acids derived from Cannabis sativa resin seizure in order to conduct a DNA typing method for the individualization of cannabis resin samples. To obtain highly purified nucleic acids from cannabis resin (Hashish) free from contaminants that cause inhibition of PCR reaction, we have tested two protocols: the CTAB protocol of Wagner and a CTAB protocol described by Somma (2004) adapted for difficult matrix. We obtained high quality genomic DNA from 8 cannabis resin seizures using the adapted protocol. DNA extracted by the Wagner CTAB protocol failed to give polymerase chain reaction (PCR) amplification of tetrahydrocannabinolic acid (THCA) synthase coding gene. However, the extracted DNA by the second protocol permits amplification of THCA synthase coding gene using different sets of primers as assessed by PCR. We describe here for the first time the possibility of DNA extraction from (Hashish) resin derived from Cannabis sativa. This allows the use of DNA molecular tests under special forensic circumstances

Prevalence of resistance to integrase strand-transfer inhibitors (INSTIs) among untreated HIV-1 infected patients in Morocco

Abstract Objective The integrase strand-transfer inhibitors (INSTIs) are an important class in the arsenal of antiretroviral drugs designed to block the integration of HIV-1 cDNA into the host DNA through the inhibition of DNA strand transfer. In this study for the first time in Morocco, the complete HIV-1 integrase gene was analysed from newly diagnosed patients to evaluate the prevalence of natural polymorphisms and INSTIs resistance-associated mutations in the integrase gene. Results The 864pb IN coding region was successfully sequenced from plasma sample for 77 among 80 antiretroviral naïve patients. The sequences were interpreted for drug resistance according to the Stanford algorithm. Sixty samples were HIV-1 subtype B (78%), fourteen CRF02_AG (18%), two subtype C and one subtype A. Overall 81 of 288 (28%) amino acid IN positions presented at least one polymorphism each. We found 18 (36.73%), 42 (25.76%) and 21 (27.27%) of polymorphic residues assigned to the N-Terminal Domain, Catalytic Core Domaine and the C-Terminal Domain positions respectively. Primary INSTIs resistance mutation were absent, however secondary mutations L74IM, T97A were detected in four samples (5.2%). These results demonstrate that untreated HIV-1 infected Moroccans will be susceptible to INSTIs

Genetic Diversity and Maternal Phylogenetic Relationships among Populations and Strains of Arabian Show Horses

Genetic diversity and phylogenetic relationships within the Arabian show horse populations are of particular interest to breeders worldwide. Using the complete mitochondrial DNA D-loop sequence (916 pb), this study aimed (i) to understand the genetic relationship between three populations, the Desert-Bred (DB), a subset of the Kingdom of Saudi Arabia (KSA), United Arab Emirates (UAE) and Bahrain (BAH), the Straight Egyptian (EG) and the Polish bloodline (PL), and (ii) to assess the accuracy of the traditional strain classification system based on maternal lines, as stated by the Bedouin culture. To that end, we collected 211 hair samples from stud farms renowned for breeding Arabian show horses from Nejd KSA, Bahrain, Egypt, Qatar, Morocco, UAE, and Poland. The phylogenetic and network analyses of the whole mitochondrial DNA D-loop sequence highlighted a great genetic diversity among the Arabian horse populations, in which about 75% of variance was assigned to populations and 25% to strains. The discriminant analysis of principal components illustrated a relative distinction between those populations. A clear subdivision between traditional strains was found in PL, in contrast to the situation of DB and EG populations. However, several Polish horse individuals could not be traced back to the Bedouin tribes by historical documentation and were shown to differ genetically from other studied Bedouin strains, hence motivating extended investigations

DNA yield and purity of isolated DNA from the eight cannabis resin samples using Wagner and adapted CTAB protocol (all DNA were re-dissolve in 100 µl sterile deionised water).

<p>DNA yield and purity of isolated DNA from the eight cannabis resin samples using Wagner and adapted CTAB protocol (all DNA were re-dissolve in 100 µl sterile deionised water).</p

(A): Agarose gel analysis (0.8%) of extracted gDNA by the adapted protocol. (B) : Agarose gel analysis (0.8%) of extracted gDNA by the Wagner protocol. (1–8): Indicated number of samples; (L): 10 kb molecular size ladder with the position of 500 bp, 1000 bp and 10 kb fragments indicated.

<p>(A): Agarose gel analysis (0.8%) of extracted gDNA by the adapted protocol. (B) : Agarose gel analysis (0.8%) of extracted gDNA by the Wagner protocol. (1–8): Indicated number of samples; (L): 10 kb molecular size ladder with the position of 500 bp, 1000 bp and 10 kb fragments indicated.</p

Sequences of oligonucleotides used in this study.

<p>Sequences of oligonucleotides used in this study.</p

Agarose gel electrophoresis (1%) of PCR amplification product of THCA synthase gene using the primers (a/b).

<p>(A): Amplification of DNA extracted by the adapted protocol. (B1): Amplification of DNA extracted by Wagner CTAB protocol (cannabis resin). (1–8): Indicated number of samples; (L): 100 bp molecular size ladder; the position of 100 bp, 600 bp and 1600 bp fragments is indicated. (B2): Amplification of DNA extracted by Wagner CTAB protocol (using other tissues of <i>cannabis sativa</i>). (GL): Grind leaves of cannabis sativa. (S): Seeds of cannabis. (R): Rod of cannabis sativa. (M): Mixed rood and leaves. (T): Negative control.</p

Example of compressed cannabis resin (Hashish) confiscated by Moroccan customs.

<p>Left hand: heated block of Hashish. Right hand: unheated block of Hashish.</p