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A BRIEF HISTORY OF GBS
Rob Elshire, Jeff Glaubitz and Katie Hyma
WHY DID WE DEVELOP GENOTYPING-BY-SEQUENCING?
Driven by the needs of the plant breeding and genetics researchers: Many markers scored quickly and cheaply.
To address the limitations of other genotyping platforms:• Cost• Throughput• Replicability• Marker Discovery Bias
GENOTYPING-BY-SEQUENCING FEATURES
• Reduced sample handling• Few PCR & purification steps• No DNA size fractionation• Efficient barcoding system• Simultaneous marker discovery &
genotyping• Scales beautifully
WHY ARE RESEARCHERS USING GBS?
• It is cheap.• Molecular biology is simple.• Produces heaps of markers.• Simultaneous discovery and genotyping.• It is robust.• Produces libraries ready to sequence on the
Illumina platform with no changes to the standard Illumina sequencing protocol or Illumina analysis pipeline.
KEYS TO GBS DATA PRODUCTION
1. High quality DNA!2. Accurate quantification. Use intercalating dye,
not spectroscopy.3. Accurate liquid handling.4. Careful titration.5. Random blank in each plate.6. Follow the protocol.
THE FUTURE OF GBS FOR MAIZE
GBS APPLIED TO MANY SPECIES AT THE INSTITUTE FOR GENOMIC DIVERSITY AT CORNELL
Barley landraces
Maize (Zea mays)
Andropogon gerardeii (big bluestem)-forage and hay
Brachypodium distachyon-model grass, biofuel crop
Rice (Oryza spp.)
Sorghum
Cucumber (cucumis sativus)
Squash (cucurbita spp.)
Soybean (Glycine max)
Potato (Solanumtuberosum)
Watermelon (Citrulluslanatus)
Pepper (capsicum annuum)
Pea (Pisum sativum)
Safflower (Carthamus tinctorius)
Hops (Humulus lupulus)-flavoring and stability in beer, herbal medicine
Chickpea (Ciser arietinum)
Sunflower (Helianthus annuus)
Horseradish (Armoraciarusticana)
Lentil (Lens culinaris)
Dioscorea (yam)
Cotton (Gopssypium spp.)
Arucariaangustifolia-seed crop, Brazil
Eucalyptus grandis-timber, pulp, riverbank stabilization
Jatropha curcas-biofuel crop
Cashew (Anacardiumoccidentale)
Willow (Salix purpurea)
Spruce (Picea spp.)
Blue catfish (Ictalurus furcatus)-Largest species of catfish in the US, primarily in the Mississippi river drainage
Gulf killfish (Fundulus grandis)-Indicator to test effects of the Deepwater Horizon oil spill
Blackstripe topminnow (Fundulusnotatus)-conservation
Mexican Tetra (blind cave fish)Astyanaz mexicanus
Verticillium dahliae(shown infecting potato tuber)
Cattle (Bos taurus) Sus scrofa (boar, pig)
Catchfly (Silene latifolia)
Neurospora spp.
Chickadee
Iris spp.
GBS STRENGTHS AND WEAKNESSES