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A rapid and hazardous reagent free protocol for genomic DNA extraction suitable for genetic studies in plants

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Abstract

Protocols for genomic DNA extraction from plants are generally lengthily, since they require that tissues be ground in liquid nitrogen, followed by a precipitation step, washing and drying of the DNA pellet, etc. This represents a major challenge especially when several hundred samples must be screened/analyzed within a working day. There is therefore a need for a rapid and simple procedure, which will produce DNA quality suitable for various analyses. Here, we describe a time and cost efficient protocol for genomic DNA isolation from plants suitable for all routine genetic screening/analyses. The protocol is free from hazardous reagents and therefore safe to be executed by non-specialists. With this protocol more than 100 genomic DNA samples could manually be extracted within a working day, making it a promising alternative in genetic studies of large-scale genomic screening projects.

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References

  1. Couch JA, Fritz PJ (1990) Isolation of DNA from plants high in polyphenolics. Plant Mol Biol Rep 8:8–12. doi:10.1007/BF02668875

    Article  CAS  Google Scholar 

  2. Ziegenhagen B, Guillemaut P, Scholz F (1993) A procedure for mini-preparation of genomic DNA from needles of silver fir (Abies alba). Plant Mol Biol Rep 11:117–121. doi:10.1007/BF02670469

    Article  CAS  Google Scholar 

  3. Chaves AL, Vergara CE, Mayer JE (1995) Dichloromethane as an economic alternative to chloroform in the extraction of DNA from plant tissues. Plant Mol Biol Rep 13:18–25. doi:10.1007/BF02668389

    Article  CAS  Google Scholar 

  4. Luro F, Laigret F (1995) Preparation of high molecular weight genomic DNA from nuclei of woody plants. Biotechniques 19:388–392

    PubMed  CAS  Google Scholar 

  5. Barnwell P, Blanchard AN, Bryant JA, Smirnoff N, Weir AF (1998) Isolation of DNA from the highly mucilaginous succulent plant Sedum telephium. Plant Mol Biol Rep 16:133–138. doi:10.1023/A:1007473302551

    Article  CAS  Google Scholar 

  6. Fang G, Hammar S, Grumet R (1992) A quick and inexpensive method for removing polysaccharides from plant genomic DNA. Biotechniques 13:52–56

    PubMed  CAS  Google Scholar 

  7. Pandey RN, Adams RP, Flournoy LE (1996) Inhibition of random amplified polymorphic DNAs (RAPDs) by plant polysaccharides. Plant Mol Biol Rep 14:17–22. doi:10.1007/BF02671898

    Article  CAS  Google Scholar 

  8. Sharma R, John SJ, Damgaard DM, McAllister TA (2003) Extraction of PCR-quality plant and microbial DNA from total rumen contents. Biotechniques 34:92–97

    PubMed  CAS  Google Scholar 

  9. Chang S, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep 11:113–116. doi:10.1007/BF02670468

    Article  CAS  Google Scholar 

  10. Sperisen C, Gugerli F, Büchler U, Mátyás G (2000) Comparison of two rapid DNA extraction protocols for gymnosperms for application in population genetic and phylogenetic studies. Genetics 7:133–136

    Google Scholar 

  11. Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 19:4321–4325. doi:10.1093/nar/8.19.4321

    Article  Google Scholar 

  12. Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW (1984) Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81:8014–8018. doi:10.1073/pnas.81.24.8014

    Article  PubMed  CAS  Google Scholar 

  13. Schmeerman MC, Mwangi J, Hobart B, Arbukle J, Vaske DA, Register JC III et al (2002) The dried corncob as a source of DNA for PCR analysis. Plant Mol Biol Rep 20:59–65. doi:10.1007/BF02801933

    Article  Google Scholar 

  14. Edwards K, Johnstone C, Thompson C (1991) A simple and rapid method for the preparation of plant genomic DNA for PCR analysis. Nucleic Acids Res 19:1349. doi:10.1093/nar/19.6.1349

    Article  PubMed  CAS  Google Scholar 

  15. Dellaporta SL, Wood J, Hicks JB (1983) Maize DNA minipreps. Maize Gent Coop News 57:26–29

    Google Scholar 

  16. Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21. doi:10.1007/BF02712670

    Article  CAS  Google Scholar 

  17. Benito CA, Figueiras M, Zaragoza C, Gallego FJ, del Pena A (1993) Rapid identification of Triticeae genotypes from single seeds using the polymerase chain reaction. Plant Mol Biol 21:181–183. doi:10.1007/BF00039629

    Article  PubMed  CAS  Google Scholar 

  18. Guidet F (1994) A powerful new technique to quickly prepare hundreds of plant extracts for PCR and RAPD analyses. Nucleic Acids Res 22:1772–1773. doi:10.1093/nar/22.9.1772

    Article  PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Agronomy Department, Purdue University, Lilly Hall, 915 West State Street, West Lafayette, IN, 47907, USA

    Simeon O. Kotchoni

  2. Department of Biology, Rutgers University, Science Building, 315 Penn Street, Camden, NJ, 08102, USA

    Emma W. Gachomo

Authors
  1. Simeon O. Kotchoni
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  2. Emma W. Gachomo
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Correspondence to Simeon O. Kotchoni.

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Kotchoni, S.O., Gachomo, E.W. A rapid and hazardous reagent free protocol for genomic DNA extraction suitable for genetic studies in plants. Mol Biol Rep 36, 1633–1636 (2009). https://doi.org/10.1007/s11033-008-9362-9

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  • DOI: https://doi.org/10.1007/s11033-008-9362-9

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