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Small-scale DNA Extraction Method for Maize and Other Plants
Published: Mar 20, 2018 DOI: 10.21769/BioProtoc.2782 Views: 7380
Reviewed by: Joëlle Schlapfer
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Abstract
Corn, or maize, is a major cereal crop and model monocot. Two of its benefits are ease of pollination and tractable genetics. For a comparison of development between maize and the dicot model, Arabidopsis, see Lunde and Hake, 2005. For an example of how this protocol might be used for a double mutant analysis see Lunde and Hake, 2009. Here, we provide a protocol for easy DNA extraction without phenol or chloroform. Therefore, this protocol is suitable for use in schools or laboratories that lack fume hoods.
Keywords: DNA extraction
Corn
Maize
Classroom
Plant Biology
Materials and Reagents
- 3.2 mm chrome beads (Bio Spec Products, catalog number: 11079132c )
- Microfuge 1.5 ml tubes (such as USA Scientific, catalog number: 1615-5500 )
- Pipette tip
- Ice
- Tris-HCl (CAS 1185-53-1)
- EDTA (CAS 6381-92-6)
- Sodium chloride (NaCl) (CAS 7647-14-5)
- Potassium acetate (KAC) (CAS 127-08-2)
- SDS (CAS 151-21-3)
- ddH2O
- Isopropanol
- 70% ethanol
- RNase (such as Sigma-Aldrich, catalog number: R6513 )
- Extraction buffer (see Recipes)
- 5 M potassium acetate (KAC) (see Recipes)
- TE with 1% RNase (see Recipes)
Equipment
- Ball Mill (such as Retsch, model: Mixer Mill MM 301 )
- QIAGEN adapter (QIAGEN, catalog number: 69982 )
- Microcentrifuge (such as Eppendorf, model: 5417 C )
- Pipette (such as Eppendorf, model: Research® plus , series in 100-1,000 μl volume)
- Vortex (such as Scientific Industries, model: Vortex-Genie 2 )
- Water bath (such as Fisher Scientific, model: Isotemp 2025 )
- pH meter (such as Thermo Fisher Scientific, Thermo ScientificTM, model: Orion StarTM A111 )
- Standard refrigerator (Such as Fisher Isotemp series)
- Spectrophotometer (such as Thermo Fisher Scientific, Thermo ScientificTM, model: NanoDropTM 1000 , catalog number: ND-1000)
- Optional equipment
- Hand drill (such as DeWalt, model: D21009 )
- Blue plastic pestle (such as DWK Life Sciences, KIMBLE, catalog number: 749521-1500 )
- Hand drill (such as DeWalt, model: D21009 )
Procedure
Category
Plant Science > Plant molecular biology > DNA
Molecular Biology > DNA > DNA extraction
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