Leaf Tissue Sampling and DNA Extraction Protocols

Taxonomists must be familiar with a number of issues in collecting and transporting samples using freezing methods (liquid nitrogen and dry ice), desiccants (silica gel and blotter paper), and preservatives (CTAB, ethanol, and isopropanol), with each method having its own merits and limitations. For most molecular studies, a reasonably good quality and quantity of DNA is required, which can only be obtained using standard DNA extraction protocols. There are many DNA extraction protocols that vary from simple and quick ones that yield low-quality DNA but good enough for routine analyses to the laborious and time-consuming standard methods that usually produce high quality and quantities of DNA. The protocol to be chosen will depend on the quality and quantity of DNA needed, the nature of samples, and the presence of natural substances that may interfere with the extraction and subsequent analysis. The protocol described in this chapter has been tested for extracting DNA from eight species and provided very good quality and quantity of DNA for different applications, including those genotyping methods that use restriction enzymes.

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Acknowledgements

This protocol was optimized for the molecular breeding component of the Drought Tolerant Maize for Africa (DTMA) project, which is funded by funded by the Bill and Melinda Gates Foundation. The author would like to thank Veronica Ogugo for testing the protocol in different species.

Author information

Authors and Affiliations

  1. International Maize and Wheat Improvement Center (CIMMYT), Nairobi, Kenya Kassa Semagn
  1. Kassa Semagn