Skip to main content

Simple, Robust Invertebrate DNA Barcoding: Chelex-Based DNA Extraction and Optimized COI Amplification

  • Protocol
  • First Online:
DNA Barcoding

Abstract

Chelex-based DNA extractions are well suited for student DNA barcoding research because they are simple, safe, and inexpensive and can be performed without specialized laboratory equipment, allowing them to be performed in classrooms or at home. Extracted DNA is stable in Chelex solution for at least a week at ambient temperature, allowing collection of DNA samples from remote students. These extractions provide quality DNA for many taxa and are optimal for barcoding invertebrates, especially in combination with novel cytochrome c oxidase I (COI) primer cocktails and PCR cycling conditions.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 219.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Hardcover Book
USD 279.99
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

Institutional subscriptions

References

  1. Hebert PD, Cywinska A, Ball SL, deWaard JR (2003) Biological identifications through DNA barcodes. Proc Biol Sci 270(1512):313–321. https://doi.org/10.1098/rspb.2002.2218

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Stoeckle M (2003) Taxonomy, DNA, and the barcode of life. Bioscience 53(9):796–797. https://doi.org/10.1641/0006-3568(2003)053[0796:TDATBC]2.0.CO;2

    Article  Google Scholar 

  3. Walsh PS, Metzger DA, Higuchi R (1991) Chelex 100 as a medium for simple extraction of DNA for PCR-based typing from forensic material. BioTechniques 10(4):506–513

    CAS  PubMed  Google Scholar 

  4. Altschmied J, Hornung U, Schlupp I, Gadau J, Kolb R, Schartl M (1997) Isolation of DNA suitable for PCR for field and laboratory work. BioTechniques 23(2):228–229. https://doi.org/10.2144/97232bm10

    Article  CAS  PubMed  Google Scholar 

  5. Gadau J (2009) DNA isolation from ants. Cold Spring Harb Protoc 2009(7):pdb.prot5245. https://doi.org/10.1101/pdb.prot5245

    Article  CAS  PubMed  Google Scholar 

  6. Micklos DA, Nash B, Hilgert U (2013) Genome science: a practical and conceptual introduction to molecular genetic analysis in eukaryotes. Cold Spring Harb Lab Press, Cold Spring Harbor, New York

    Google Scholar 

  7. Hyman OJ, Doyle EA, Harsh J, Mott J, Pesce A, Rasoul B, Seifert K, Enke RA (2019) CURE-all: large scale implementation of authentic DNA barcoding research into first-year biology curriculum. CourseSource. https://doi.org/10.24918/cs.2019.10

  8. Marizzi C, Florio A, Lee M, Khalfan M, Ghiban C, Nash B, Dorey J, McKenzie S, Mazza C, Cellini F, Baria C, Bepat R, Cosentino L, Dvorak A, Gacevic A, Guzman-Moumtzis C, Heller F, Holt NA, Horenstein J, Joralemon V, Kaur M, Kaur T, Khan A, Kuppan J, Laverty S, Lock C, Pena M, Petrychyn I, Puthenkalam I, Ram D, Ramos A, Scoca N, Sin R, Gonzalez I, Thakur A, Usmanov H, Han K, Wu A, Zhu T, Micklos DA (2018) DNA barcoding Brooklyn (New York): a first assessment of biodiversity in Marine Park by citizen scientists. PLoS One 13(7):e0199015. https://doi.org/10.1371/journal.pone.0199015

    Article  PubMed  PubMed Central  Google Scholar 

  9. Pepenella S, Hackett J, Fernandez-Marco C, Petracca J, Marizzi C, Nash B, Micklos DA (2024) Rapid, equipment-free DNA isolation on chromatography paper for DNA barcoding. In: Desalle R (ed) DNA barcoding, methods and protocols, Methods in molecular biology. Springer, New York

    Google Scholar 

  10. Folmer O, Black M, Hoeh W, Lutz R, Vrijenhoek R (1994) DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Mol Mar Biol Biotechnol 3(5):294–299

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Jennifer Hackett .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Hackett, J. et al. (2024). Simple, Robust Invertebrate DNA Barcoding: Chelex-Based DNA Extraction and Optimized COI Amplification. In: DeSalle, R. (eds) DNA Barcoding. Methods in Molecular Biology, vol 2744. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3581-0_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-3581-0_6

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3580-3

  • Online ISBN: 978-1-0716-3581-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics