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MB2 - Selection of Markers for Molecular Breeding

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DNA extraction protocols

Which DNA extraction protocol you decide to use depends on the quality and quantity of DNA needed, the type of assay you want to utilize, and the number of markers needed.  Here is a list of 3 common protocols and each of their advantages:

  1. CTAB (cetyltrimethyl ammonium bromide), large prep, miniprep: very clean DNA, large quantity, long process, and may use potentially dangerous chemicals such as chloroform, as seen in Figure 1. (see some examples here: http://www.protocol-online.org/prot/Molecular_Biology/DNA/DNA_Extraction___Purification/DNA_Extraction_from_Plants/index.html
  2. Whatman FTA cards (fast technology for analysis of nucleic acids): less clean, small quantity, fast and easy: http://www.gelifesciences.com/webapp/wcs/stores/servlet/catalog/en/GELifeSciences-us/products/AlternativeProductStructure_17096/
  3. Qiagen column kits: very clean, good quantity, fairly simple: http://www.qiagen.com/products/catalog/sample-technologies/dna-sample-technologies/genomic-dna/dneasy-plant-mini-kit#  (qiagen.com )
  4. Many others…..

Figure 3 shows an example of the FTA card, where liquid from a leaf sample is eluted on to the card, and a punched disk is used to extract DNA and an example of a Qiagen kit, with all the ingredients needed to extract DNA from a leaf sample included. 

fta card.jpgqiagen.jpg

Figure 3 Top:  Examples of DNA extraction protocol kits.

Images taken from company websites as previously mentioned.

 

 

Figure 3 Bottom:
Example of a Qiagen kit


For a more indepth discussion and/or review of DNA in general and how the extraction process works, please refer to eLesson, “DNA and DNA Extraction” by Patricia Hain and Donald Lee.

 

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