Plant Molecular Biology Reporter

, Volume 21, Issue 4, pp 459–460

A high-throughput DNA extraction protocol for tropical molecular breeding programs

  • Emma S. Mace
  • Kutokshi K. Buhariwalla
  • Hutokshi K. Buhariwalla
  • Jonathan H. Crouch
Publish by Abstract

DOI: 10.1007/BF02772596

Cite this article as:
Mace, E.S., Buhariwalla, K.K., Buhariwalla, H.K. et al. Plant Mol Biol Rep (2003) 21: 459. doi:10.1007/BF02772596

Abstract

Liquid handling robotics and capillary electrophoresis genetic analyzers now offer high-throughput solutions for 2 of the 4 key steps in PCR-based DNA marker-assisted fingerprinting (DNA extraction, PCR amplification, electrophoresis, data analysis). Thus, DNA extraction remains the most significant bottleneck at the bench for large-scale applications in plant breeding and germplasm characterization. We report on a rapid and low-cost method for relatively high-throughput extraction of high-quality DNA from young and mature leaves of sorghum, pearl millet, chickpea, groundnut, and pigeonpea. The procedure uses a modified CTAB/β-mercaptoethanol method for DNA extraction in a 96-well plate. The quantity and quality of the DNA extracted per sample is adequate for more than 1000 PCR reactions. A relatively high throughput of 96–384 samples per person per day can be achieved, depending on the crop. A major timesaving aspect of the protocol is the absence of a manual sample-grinding step. Finally, the cost is a magnitude lower than commercial plate-based kits, and, as such, is likely to have substantial application in tropical molecular breeding programs.

Key words

DNA isolation high throughput molecular breeding PCR 

Copyright information

© International Society for Plant Molecular Biology 2003

Authors and Affiliations

  • Emma S. Mace
    • 2
  • Kutokshi K. Buhariwalla
    • 2
  • Hutokshi K. Buhariwalla
    • 2
  • Jonathan H. Crouch
    • 2
  1. 1.MS Swaminathan Applied Genomics LaboratoryICRISAT (International Crops Research Institute for the Semi-Arid Tropics)PatancheruIndia
  2. 2.Hermitage Research StationWarwickAustralia

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