Skip to main content

Capillary Electrophoresis of DNA from Cannabis sativa for Correlation of Samples to Geographic Origin

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 830))

Abstract

For routine genetic analysis of Cannabis sativa, two methods are currently in use, (a) AFLP; amplified fragment length polymorphism analysis and (b) STR; short tandem repeat analysis. The AFLP method used on capillary electrophoresis instrumentation is fully described in this chapter. AFLP analysis generates numerous nonspecific marker fragments for a complex DNA pattern and is available in kit format for quality assurance of reagents. This method is particularly useful when discerning the genetics of highly inbred plant species that may share much of the same DNA with only slight differences due to their common genetic background. AFLP analysis, however, is most effective on fresh or well-preserved plant specimens where the integrity of the DNA is high and the sample is a single source specimen (i.e., not a mixture of plants or different species).

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   159.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Office of National Drug Control Policy (2007) Marijuana on public lands. http://White-HouseDrugPolicy.gov. Accessed 07 January 2011

  2. Miller Coyle H, Palmbach T, Juliano N, Ladd C, Lee HC (2003) An overview of DNA methods for the identification and individualization of marijuana. Croat Med J 44(3):315–321

    PubMed  Google Scholar 

  3. Howard C, Gilmore S, Robertson J, Peakall R (2009) A Cannabis sativa STR genotype database for Australian seizures: forensic applications and limitations. J Forensic Sci 54(3):556–563

    Article  PubMed  CAS  Google Scholar 

  4. Alghanim HJ, Almirall JR (2003) Development of microsatellite markers in Cannabis sativa for DNA typing and genetic relatedness analyses. Anal Bioanal Chem 376(8):1225–1233

    Article  PubMed  CAS  Google Scholar 

  5. Hurley JM, West JB, Ehleringer JR (2010) Stable isotope models to predict geographic origin and cultivation conditions of marijuana. Sci Justice 50(2):86–93

    Article  PubMed  CAS  Google Scholar 

  6. Fry NK, Savelkoul PH, Visca P (2009) Amplified fragment-length polymorphism analysis. Methods Mol Biol 551:89–104

    Article  PubMed  CAS  Google Scholar 

  7. Gaggiotti OE (2010) Bayesian statistical treatment of the fluorescence of AFLP bands leads to accurate genetic structure inference. Mol Ecol 19(21):4586–4588

    Article  PubMed  CAS  Google Scholar 

  8. Howard C, Gilmore S, Robertson J, Peakall R (2008) Developmental validation of a Cannabis sativa STR multiplex system for forensic analysis. J Forensic Sci 53(5):1061–1067

    PubMed  CAS  Google Scholar 

  9. Hsieh HM, Hou RJ, Tsai LC, Wei CS, Liu SW, Huang LH, Kuo YC, Linacre A, Lee JC (2003) A highly polymorphic STR locus in Cannabis sativa. Forensic Sci Int 131(1):53–58

    Article  PubMed  CAS  Google Scholar 

  10. Mendoza MA, Mills DK, Lata H, Chandra S, ElSohly MA, Almirall JR (2009) Genetic individualization of Cannabis sativa by a short tandem repeat multiplex system. Anal Bioanal Chem 393(2):719–726

    Article  PubMed  CAS  Google Scholar 

  11. Gilmore S, Peakall R, Robertson J (2003) Short tandem repeat (STR) DNA markers are hypervariable and informative in Cannabis sativa: implications for forensic investigations. Forensic Sci Int 131(1):65–67

    Article  PubMed  CAS  Google Scholar 

  12. Ergül A, Kazan K, Aras S, Cevik V, Celik H, Söylemezoğlu G (2006) AFLP analysis of genetic variation within the two economically important Anatolian grapevine (Vitis vinifera L.) varietal groups. Genome 49(5):467–475

    Article  PubMed  Google Scholar 

  13. Jakse J, Kindlhofer K, Javornik B (2001) Assessment of genetic variation and differentiation of hop genotypes by microsatellite and AFLP markers. Genome 44(5):773–782

    Article  PubMed  CAS  Google Scholar 

  14. Kuhlman LC, Burson BL, Stelly DM, Klein PE, Klein RR, Price HJ, Rooney WL (2010) Early-generation germplasm introgression from Sorghum macrospermum into sorghum (S. bicolor). Genome 53(6):419–429

    Article  PubMed  CAS  Google Scholar 

  15. Opara UL, Jacobson D, Al-Saady NA (2010) Analysis of genetic diversity in banana cultivars (Musa cvs.) from the South of Oman using AFLP markers and classification by phylogenetic, hierarchical clustering and principal component analyses. J Zhejiang Univ Sci B 11(5):332–341

    Article  PubMed  CAS  Google Scholar 

  16. Rahman MH, Rajora OP (2002) Microsatellite DNA fingerprinting, differentiation, and genetic relationships of clones, cultivars, and varieties of six poplar species from three sections of the genus Populus. Genome 45(6):1083–1094

    Article  PubMed  CAS  Google Scholar 

  17. Rajora P, Rahman H (2003) Microsatellite DNA and RAPD fingerprinting, identification and genetic relationships of hybrid poplar (Populus x canadensis) cultivars. Theor Appl Genet 106(3):470–477

    PubMed  CAS  Google Scholar 

  18. Stefanowicz-Hajduk J, Filipowicz N, Kosinski I, Ochocka JR (2009) Capillary electrophoresis with dual laser detection in separation of amplified fragment length polymorphism fragments. J Sep Sci 32(20):3539–3543

    Article  PubMed  CAS  Google Scholar 

  19. (2010) AFLP Plant Mapping Protocol. Applied Biosystems, Inc. Foster City, CA

    Google Scholar 

  20. Miller Coyle H (2005) Forensic botany: principles and applications to criminal casework. CRC Press, Boca Raton, FL

    Google Scholar 

  21. Miller Coyle H, Shutler G, Tully L, Pagliaro E, Harper A, Palmbach T, Lee HC (2005) Validation of a DNA method for the individualization of plant evidence. http://www.ncjrs.gov/pdffiles1/nij/grants/211999pdf. Accessed 08 January 2011

Download references

Acknowledgments

This work was previously supported by the National Institute of Justice Grant #2001-IJ-CX-K011. Thank you to the Connecticut State Forensic Science Laboratory and the University of New Haven for use of their facilities. Numerous students, scientists, and task force personnel offered advice or generated data sets during the course of the grant period. Thank you to Dr. Henry Lee, Major Timothy Palmbach, Dr. Eric Buel, Captain Peter Warren, members of the Connecticut Statewide Narcotics Task Force, Eric Carita, Nicholas C.S. Yang, Elizabeth McClure Baker, Joselle Germano, Edward Jachimowicz III, Shane Lumpkins, Khartika Divarkaran, and Michelle Irvin for their work on the grant funded project.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Heather Miller Coyle .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Coyle, H.M. (2012). Capillary Electrophoresis of DNA from Cannabis sativa for Correlation of Samples to Geographic Origin. In: Alonso, A. (eds) DNA Electrophoresis Protocols for Forensic Genetics. Methods in Molecular Biology, vol 830. Humana Press. https://doi.org/10.1007/978-1-61779-461-2_17

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-461-2_17

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-460-5

  • Online ISBN: 978-1-61779-461-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics