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Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing

BACKGROUND: Marijuana (Cannabis sativa) is the most commonly used illicit drug in the USA, and the use of DNA barcodes could assist drug trafficking investigations by indicating the biogeographical origin and crop type of a sample and providing a means for linking cases. Additionally, the legality o...

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Autores principales: Roman, Madeline G., Gutierrez, Ryan, Houston, Rachel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8928601/
https://www.ncbi.nlm.nih.gov/pubmed/35300721
http://dx.doi.org/10.1186/s42238-022-00123-2
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author Roman, Madeline G.
Gutierrez, Ryan
Houston, Rachel
author_facet Roman, Madeline G.
Gutierrez, Ryan
Houston, Rachel
author_sort Roman, Madeline G.
collection PubMed
description BACKGROUND: Marijuana (Cannabis sativa) is the most commonly used illicit drug in the USA, and the use of DNA barcodes could assist drug trafficking investigations by indicating the biogeographical origin and crop type of a sample and providing a means for linking cases. Additionally, the legality of marijuana in the USA remains complicated with some states fully legalizing marijuana for recreational use while federally marijuana remains completely illegal. Massively parallel sequencing (MPS) offers distinct advantages over capillary electrophoresis (CE), including more comprehensive coverage of target loci, analysis of hundreds of markers simultaneously, and high throughput capabilities. METHODS: This study reports on the development of a MiSeq FGx® assay targeting seven “hotspot” regions in the Cannabis sativa chloroplast genome that are highly polymorphic and informative in attempts to determine biogeographical origin and distinguishing between marijuana and hemp. Sequencing results were compared to previous studies that used CE-based genotyping methods. RESULTS: A total of 49 polymorphisms were observed, 16 of which have not been previously reported. Additionally, sequence data revealed isoalleles at one locus, which were able to differentiate two samples that had the same haplotype using CE-based methods. This study reports preliminary results from sequencing 14 hemp and marijuana samples from different countries using the developed MPS assay. CONCLUSION: Future studies should genotype a more comprehensive sample set from around the world to build a haplotype database, which could be used to provide investigative leads for law enforcement agencies investigating marijuana trafficking. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s42238-022-00123-2.
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spelling pubmed-89286012022-03-23 Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing Roman, Madeline G. Gutierrez, Ryan Houston, Rachel J Cannabis Res Brief Research Report BACKGROUND: Marijuana (Cannabis sativa) is the most commonly used illicit drug in the USA, and the use of DNA barcodes could assist drug trafficking investigations by indicating the biogeographical origin and crop type of a sample and providing a means for linking cases. Additionally, the legality of marijuana in the USA remains complicated with some states fully legalizing marijuana for recreational use while federally marijuana remains completely illegal. Massively parallel sequencing (MPS) offers distinct advantages over capillary electrophoresis (CE), including more comprehensive coverage of target loci, analysis of hundreds of markers simultaneously, and high throughput capabilities. METHODS: This study reports on the development of a MiSeq FGx® assay targeting seven “hotspot” regions in the Cannabis sativa chloroplast genome that are highly polymorphic and informative in attempts to determine biogeographical origin and distinguishing between marijuana and hemp. Sequencing results were compared to previous studies that used CE-based genotyping methods. RESULTS: A total of 49 polymorphisms were observed, 16 of which have not been previously reported. Additionally, sequence data revealed isoalleles at one locus, which were able to differentiate two samples that had the same haplotype using CE-based methods. This study reports preliminary results from sequencing 14 hemp and marijuana samples from different countries using the developed MPS assay. CONCLUSION: Future studies should genotype a more comprehensive sample set from around the world to build a haplotype database, which could be used to provide investigative leads for law enforcement agencies investigating marijuana trafficking. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s42238-022-00123-2. BioMed Central 2022-03-17 /pmc/articles/PMC8928601/ /pubmed/35300721 http://dx.doi.org/10.1186/s42238-022-00123-2 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Brief Research Report
Roman, Madeline G.
Gutierrez, Ryan
Houston, Rachel
Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing
title Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing
title_full Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing
title_fullStr Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing
title_full_unstemmed Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing
title_short Massively parallel sequencing of Cannabis sativa chloroplast hotspots for forensic typing
title_sort massively parallel sequencing of cannabis sativa chloroplast hotspots for forensic typing
topic Brief Research Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8928601/
https://www.ncbi.nlm.nih.gov/pubmed/35300721
http://dx.doi.org/10.1186/s42238-022-00123-2
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